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<meta name="description" content="Tunable control of gene expression is an invaluable tool for biological experiments. In this study, we describe a new xylose-inducible promoter system and evaluate it in both &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; and &lt;i&gt;Pseudomonas fluorescens&lt;/i&gt; The &lt;i&gt;P&lt;sub&gt;xut&lt;/sub&gt;&lt;/i&gt; promoter, derived from the &lt;i&gt;P.&lt;/i&gt; …"><meta name="keywords" content="pmid:32414795, PMC7357479, doi:10.1128/AEM.00250-20, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov&#x27;t, Jake D Callaghan, Nicholas A Stella, Robert M Q Shanks, Bacterial Proteins / genetics*, Bacterial Proteins / metabolism, Base Sequence, Epithelium, Corneal / injuries, Membrane Proteins / genetics*, Membrane Proteins / metabolism, Promoter Regions, Genetic*, Pseudomonas aeruginosa / genetics*, Pseudomonas aeruginosa / metabolism, Pseudomonas fluorescens / genetics*, Pseudomonas fluorescens / metabolism, Re-Epithelialization / genetics*, Sequence Analysis, DNA, Type II Secretion Systems / genetics*, Type II Secretion Systems / metabolism, Xylose / metabolism*, PubMed Abstract, NIH, NLM, NCBI, National Institutes of Health, National Center for Biotechnology Information, National Library of Medicine, MEDLINE"><meta name="robots" content="index,nofollow,noarchive"><meta property="og:title" content="Xylose-Inducible Promoter Tools for Pseudomonas Species and Their Use in Implicating a Role for the Type II Secretion System Protein XcpQ in the Inhibition of Corneal Epithelial Wound Closure - PubMed"><meta property="og:url" content="https://pubmed.ncbi.nlm.nih.gov/32414795/"><meta property="og:description" content="Tunable control of gene expression is an invaluable tool for biological experiments. In this study, we describe a new xylose-inducible promoter system and evaluate it in both &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; and &lt;i&gt;Pseudomonas fluorescens&lt;/i&gt; The &lt;i&gt;P&lt;sub&gt;xut&lt;/sub&gt;&lt;/i&gt; promoter, derived from the &lt;i&gt;P.&lt;/i&gt; …"><meta property="og:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:image:secure_url" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:type" content="website"><meta property="og:site_name" content="PubMed"><meta name="twitter:domain" content="pubmed.ncbi.nlm.nih.gov"><meta name="twitter:card" content="summary_large_image"><meta name="twitter:title" content="Xylose-Inducible Promoter Tools for Pseudomonas Species and Their Use in Implicating a Role for the Type II Secretion System Protein XcpQ in the Inhibition of Corneal Epithelial Wound Closure - PubMed"><meta name="twitter:url" content="https://pubmed.ncbi.nlm.nih.gov/32414795/"><meta name="twitter:description" content="Tunable control of gene expression is an invaluable tool for biological experiments. In this study, we describe a new xylose-inducible promoter system and evaluate it in both &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; and &lt;i&gt;Pseudomonas fluorescens&lt;/i&gt; The &lt;i&gt;P&lt;sub&gt;xut&lt;/sub&gt;&lt;/i&gt; promoter, derived from the &lt;i&gt;P.&lt;/i&gt; …"><meta name="twitter:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg">
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<span class="email-subject" id="email-subject" aria-label="Email subject">1 selected item: 32414795 - PubMed</span>
<input type="hidden" name="email-subject" value="1 selected item: 32414795"/>
<input type="hidden" name="email-subject-hash" value="daf73f4f9a2acef739f043ac78057b6bc061a701785f56c02df713b543de1d78"/>
</div>
<div class="action-panel-control-wrap">
<label for="email-to" class="action-panel-label required-field-asterisk">
To:
</label>
<input type="email" aria-label="Recipient Email Address" placeholder="email@example.com" id="email-to" class="email-to" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="100" required>
</div>
<div class="action-panel-control-wrap">
<label for="email-from" class="action-panel-label">
From:
</label>
<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
</div>
<div class="action-panel-control-wrap">
<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="rXZRnk5Bm5U2iDXt4luaCkISePYHWdmkIKJW0ayXWGHwc43jHE7c6sewMTGTPOUE">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
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<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="rXZRnk5Bm5U2iDXt4luaCkISePYHWdmkIKJW0ayXWGHwc43jHE7c6sewMTGTPOUE">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
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<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
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<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="rXZRnk5Bm5U2iDXt4luaCkISePYHWdmkIKJW0ayXWGHwc43jHE7c6sewMTGTPOUE">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
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<a class="link-item
dialog-focus"
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Full text links
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<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/32414795/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/32414795/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="32414795" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F32414795%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="rXZRnk5Bm5U2iDXt4luaCkISePYHWdmkIKJW0ayXWGHwc43jHE7c6sewMTGTPOUE"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
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Xylose-Inducible Promoter Tools for <i>Pseudomonas</i> Species and Their Use in Implicating a Role for the Type II Secretion System Protein XcpQ in the Inhibition of Corneal Epithelial Wound Closure
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Xylose-Inducible Promoter Tools for <i>Pseudomonas</i> Species and Their Use in Implicating a Role for the Type II Secretion System Protein XcpQ in the Inhibition of Corneal Epithelial Wound Closure
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><sup class="key">1</sup> Charles T. Campbell Laboratory of Ophthalmic Microbiology, Department of Ophthalmology, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania, USA.</li>
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><sup class="key">2</sup> Department of Ophthalmology, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania, USA.</li>
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><sup class="key">3</sup> Charles T. Campbell Laboratory of Ophthalmic Microbiology, Department of Ophthalmology, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania, USA shanksrm@upmc.edu.</li>
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Abstract
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<p>
Tunable control of gene expression is an invaluable tool for biological experiments. In this study, we describe a new xylose-inducible promoter system and evaluate it in both <i>Pseudomonas aeruginosa</i> and <i>Pseudomonas fluorescens</i> The <i>P<sub>xut</sub></i> promoter, derived from the <i>P. fluorescens</i><i>xut</i> operon, was incorporated into a broad-host-range pBBR1-based plasmid and was compared to the <i>Escherichia coli</i>-derived <i>P<sub>BAD</sub></i> promoter using <i>gfp</i> as a reporter. Green fluorescent protein (GFP) fluorescence from the <i>P<sub>xut</sub></i> promoter was inducible in both <i>Pseudomonas</i> species, but not in <i>E. coli</i>, which may facilitate the cloning of genes toxic to <i>E. coli</i> to generate plasmids. The <i>P<sub>xut</sub></i> promoter was activated at a lower inducer concentration than <i>P<sub>BAD</sub></i> in <i>P. fluorescens</i>, and higher <i>gfp</i> levels were achieved using <i>P<sub>xut</sub></i> Flow cytometry analysis indicated that <i>P<sub>xut</sub></i> was leakier than <i>P<sub>BAD</sub></i> in the <i>Pseudomonas</i> species tested but was expressed in a higher proportion of cells when induced. d-Xylose as a sole carbon source did not support the growth of <i>P. aeruginosa</i> or <i>P. fluorescens</i> and is less expensive than many other commonly used inducers, which could facilitate large-scale applications. The efficacy of this system was demonstrated by its use to reveal a role for the <i>P. aeruginosa</i> type II secretion system gene <i>xcpQ</i> in bacterial inhibition of corneal epithelial cell wound closure. This study introduces a new inducible promoter system for gene expression for use in <i>Pseudomonas</i> species.<b>IMPORTANCE</b><i>Pseudomonas</i> species are enormously important in human infections, in biotechnology, and as model systems for investigating basic science questions. In this study, we have developed a xylose-inducible promoter system, evaluated it in <i>P. aeruginosa</i> and <i>P. fluorescens</i>, and found it to be suitable for the strong induction of gene expression. Furthermore, we have demonstrated its efficacy in controlled gene expression to show that a type II secretion system protein from <i>P. aeruginosa</i>, XcpQ, is important for host-pathogen interactions in a corneal wound closure model.
</p>
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<p>
<strong class="sub-title">
Keywords:
</strong>
Pseudomonas; inducible promoter; plasmids; xylose.
</p>
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Copyright © 2020 American Society for Microbiology.
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<figure class="figure-item "
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data-slide-index="0"
data-label-slug="fig-1">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/b7a3323a87d7/AEM.00250-20-f0001.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-0"
role="button"
data-image-width="760"
data-image-height="497"
data-image-alt="FIG 1"
data-pmc-id="PMC7357479"
data-figure-id="F1">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/df50f605f557/AEM.00250-20-f0001.gif"
alt="FIG 1" />
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<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 1 </p>
</strong>
<div class="figure-caption-contents"><p> Xylose repressor genomic region from… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 1 </p>
</strong>
<div class="figure-caption-contents"><p> Xylose repressor genomic region from P. fluorescens strain Pf0-1. (A) Genetic map of… </p></div>
</div>
<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 1
</strong>
<div class="figure-caption-contents">Xylose repressor genomic region from P. fluorescens strain Pf0-1. (A) Genetic map of the <i>xutR-xutA</i> region. The shaded bar represents the 188-bp intergenic region. (B) DNA sequence of the <i>P<sub>xut</sub></i> promoter upstream of Pf101_2303. The conserved operator inverted repeats are shown in boldface, and the 35 and 10 regions for the <i>xutA</i> promoter (<i>P<sub>xut</sub></i>) are underlined. A direct repeat of 13 bp is shown in italics. A putative ribosome binding site 9 to 12 bp upstream of the start codon is shown in lowercase. (C) Schematic diagrams of select plasmids used in this study.</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="1"
data-label-slug="fig-2">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/a63b3484142a/AEM.00250-20-f0002.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-1"
role="button"
data-image-width="750"
data-image-height="248"
data-image-alt="FIG 2"
data-pmc-id="PMC7357479"
data-figure-id="F2">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/04675b3294cc/AEM.00250-20-f0002.gif"
alt="FIG 2" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="750">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="248">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 2 </p>
</strong>
<div class="figure-caption-contents"><p> Comparison of the <i> P <sub> BAD… </sub> </i> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 2 </p>
</strong>
<div class="figure-caption-contents"><p> Comparison of the <i> P <sub> BAD </sub> </i> and <i> P <sub> xut </sub> </i> promoters in E. coli and… </p></div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 2
</strong>
<div class="figure-caption-contents">Comparison of the <i>P<sub>BAD</sub></i> and <i>P<sub>xut</sub></i> promoters in E. coli and <i>Pseudomonas</i> species. Shown is the expression of <i>gfpmut3</i> from the two different promoters in a pBBR1-based plasmid. Cultures were prepared in LB medium with a range of inducer concentrations and were grown at 30°C for 25 h. Means and standard deviations are shown (<i>n</i>, ≥5 independent cultures). <i>P<sub>xut</sub></i> was not inducible in E. coli and was inducible at lower inducer concentrations than <i>P<sub>BAD</sub></i> in P. fluorescens. The pMQ578 plasmid has <i>P<sub>xut</sub>-gfp</i>, and pMQ80 has <i>P<sub>BAD</sub>-gfp</i>. Asterisks indicate statistically significant differences (<i>P</i>&lt;0.05) between groups by Students <i>t</i> test.</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="2"
data-label-slug="fig-3">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/33470f3ac923/AEM.00250-20-f0003.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-2"
role="button"
data-image-width="750"
data-image-height="280"
data-image-alt="FIG 3"
data-pmc-id="PMC7357479"
data-figure-id="F3">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/9c7f17017d20/AEM.00250-20-f0003.gif"
alt="FIG 3" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="750">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="280">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 3 </p>
</strong>
<div class="figure-caption-contents"><p> Expression of the <i> P <sub> BAD… </sub> </i> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 3 </p>
</strong>
<div class="figure-caption-contents"><p> Expression of the <i> P <sub> BAD </sub> </i> and <i> P <sub> xut </sub> </i> promoters in <i> Pseudomonas </i> species and… </p></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 3
</strong>
<div class="figure-caption-contents">Expression of the <i>P<sub>BAD</sub></i> and <i>P<sub>xut</sub></i> promoters in <i>Pseudomonas</i> species and E. coli over time with 10mM inducer. Shown is the expression of <i>gfpmut3</i> over time from the two different promoters in a pBBR1-based plasmid. Cultures were prepared in LB medium with an inducer (10mM) and were grown at 30°C. Means and standard deviations are shown (<i>n</i>, ≥4 independent cultures for E. coli; <i>n</i>, 6 for <i>Pseudomonas</i> species). The pMQ578 plasmid has <i>P<sub>xut</sub>-gfp</i>, and pMQ80 has <i>P<sub>BAD</sub>-gfp</i>. Xylose was used as an inducer for pMQ578, and arabinose was used for pMQ80. Asterisks indicate statistically significant differences (<i>P</i>&lt;0.05) between groups by Students <i>t</i> test.</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="3"
data-label-slug="fig-4">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/7ce1fec0e9a6/AEM.00250-20-f0004.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="750"
data-image-height="399"
data-image-alt="FIG 4"
data-pmc-id="PMC7357479"
data-figure-id="F4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/fed6dc95fea4/AEM.00250-20-f0004.gif"
alt="FIG 4" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="750">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="399">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 4 </p>
</strong>
<div class="figure-caption-contents"><p> The <i> Pseudomonas </i> species tested do… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 4 </p>
</strong>
<div class="figure-caption-contents"><p> The <i> Pseudomonas </i> species tested do not use <sc> d </sc> -xylose as a sole carbon… </p></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 4
</strong>
<div class="figure-caption-contents">The <i>Pseudomonas</i> species tested do not use <sc>d</sc>-xylose as a sole carbon source. Cultures were incubated in M9 minimal medium with the indicated sugars at 10mM for 25 hat 30°C. Means and standard deviations are shown (<i>n</i>, 6 independent cultures). Asterisks indicate significant differences from the glucose group (<i>P</i>&lt;0.05) by ANOVA with Tukeys posttest.</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="4"
data-label-slug="fig-5">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/394f5d980528/AEM.00250-20-f0005.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="742"
data-image-height="358"
data-image-alt="FIG 5"
data-pmc-id="PMC7357479"
data-figure-id="F5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/be8e3721a3cf/AEM.00250-20-f0005.gif"
alt="FIG 5" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="742">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="358">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 5 </p>
</strong>
<div class="figure-caption-contents"><p> Evaluation of ribose for induction… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 5 </p>
</strong>
<div class="figure-caption-contents"><p> Evaluation of ribose for induction of the <i> P <sub> xut </sub> </i> promoter in two <i> Pseudomonas… </i> </p></div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 5
</strong>
<div class="figure-caption-contents">Evaluation of ribose for induction of the <i>P<sub>xut</sub></i> promoter in two <i>Pseudomonas</i> species. Shown is the expression of <i>gfpmut3</i> in a pBBR1-based plasmid. Cultures were prepared in LB medium with a range of inducer concentrations and were grown at 30°C for 25 h. Means and standard deviations are given (<i>n</i>, 6 independent cultures). (A) <i>P<sub>xut</sub></i>-based GFP expression was minimally induced by different ribose concentrations. Asterisks indicate significant differences compared to the no-ribose group as determined by Student's <i>t</i> test (<i>P</i> &lt; 0.05). (B) Ribose (10 mM) did not alter the ability of xylose (10 mM) to activate <i>P<sub>xut</sub></i>-based GFP expression in P. aeruginosa strain PA14. Asterisks indicate significant differences compared to the water group as determined by ANOVA with Tukey's posttest (<i>P</i> &lt; 0.05).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="5"
data-label-slug="fig-6">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/082bde700c36/AEM.00250-20-f0006.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="680"
data-image-height="562"
data-image-alt="FIG 6"
data-pmc-id="PMC7357479"
data-figure-id="F6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/95577b660805/AEM.00250-20-f0006.gif"
alt="FIG 6" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="680">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="562">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 6 </p>
</strong>
<div class="figure-caption-contents"><p> Stationary-phase induction of the <i> P… </i> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 6 </p>
</strong>
<div class="figure-caption-contents"><p> Stationary-phase induction of the <i> P <sub> BAD </sub> </i> and <i> P <sub> xut </sub> </i> promoters in <i> Pseudomonas </i> species.… </p></div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 6
</strong>
<div class="figure-caption-contents">Stationary-phase induction of the <i>P<sub>BAD</sub></i> and <i>P<sub>xut</sub></i> promoters in <i>Pseudomonas</i> species. Bacteria were grown overnight in LB broth, washed, and adjusted to an OD<sub>600</sub> of 2.0 in PBS with an inducer at 10mM. GFP fluorescence was measured over time. Means and standard deviations are given (<i>n</i>, 3 to 4 independent cultures). <i>P<sub>xut</sub></i> was more highly inducible in P. fluorescens than in P. aeruginosa during stationary phase. <i>P<sub>xut</sub></i> and <i>P<sub>BAD</sub></i> were indistinguishable in P. aeruginosa. The pMQ578 plasmid has <i>P<sub>xut</sub>-gfp</i>, and pMQ80 has <i>P<sub>BAD</sub>-gfp</i>. The asterisk indicates a statistically significant difference (<i>P</i>&lt;0.05) by Students <i>t</i> test between normalized fluorescence levels for Pf0-1 with different plasmids; there were no differences between plasmids in fluorescence levels for PA14. At 25 h, Pf0-1 with pMQ578 produced significantly more fluorescence than PA14.</div>
</figcaption>
</figure>
<figure class="figure-item tail"
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itemprop="contentUrl"
aria-describedby="figure-caption-6"
role="button"
data-image-width="742"
data-image-height="742"
data-image-alt="FIG 7"
data-pmc-id="PMC7357479"
data-figure-id="F7">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-6"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/81998caba9e3/AEM.00250-20-f0007.gif"
alt="FIG 7" />
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<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 7 </p>
</strong>
<div class="figure-caption-contents"><p> Flow cytometry analysis of <i> P… </i> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 7 </p>
</strong>
<div class="figure-caption-contents"><p> Flow cytometry analysis of <i> P <sub> BAD </sub> </i> and <i> P <sub> xut </sub> </i> promoter-driven GFP expression in… </p></div>
</div>
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 7
</strong>
<div class="figure-caption-contents">Flow cytometry analysis of <i>P<sub>BAD</sub></i> and <i>P<sub>xut</sub></i> promoter-driven GFP expression in <i>Pseudomonas</i> species indicates increased leakiness and increased proportions of bacteria expressing GFP with an inducer. The expression of <i>gfpmut3</i> from the two different promoters was measured by flow cytometry. Cultures were prepared in LB medium with an inducer (10mM) and were grown at 30°C. Gray areas represent PA14 or Pf0-1 with no plasmid, used to determine background levels. Yellow peaks represent bacteria with pMQ578 or pMQ80 without an inducer, used to indicate leakiness. Blue (and green) peaks represent bacteria with pMQ578 or pMQ80 and with an inducer. The pMQ578 plasmid has <i>P<sub>xut</sub>-gfp</i>, and pMQ80 has <i>P<sub>BAD</sub>-gfp</i>. Xylose was used as an inducer for pMQ578, and arabinose was used for pMQ80. Levels of GFP fluorescence above those for bacteria without a plasmid were used to determine the positive cutoff (noted as GFP+); horizontal green bars indicate positive values. Results of a representative experiment are shown (<i>n</i>, 100,000 cells per group).</div>
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itemprop="contentUrl"
aria-describedby="figure-caption-7"
role="button"
data-image-width="750"
data-image-height="406"
data-image-alt="FIG 8"
data-pmc-id="PMC7357479"
data-figure-id="F8">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-7"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/148c3e1ddc58/AEM.00250-20-f0008.gif"
alt="FIG 8" />
</a>
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<strong class="figure-label">
<p> FIG 8 </p>
</strong>
<div class="figure-caption-contents"><p> Analysis of promoter leakiness and… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 8 </p>
</strong>
<div class="figure-caption-contents"><p> Analysis of promoter leakiness and strength using flow cytometry. GFP expression in <i> Pseudomonas… </i> </p></div>
</div>
<figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 8
</strong>
<div class="figure-caption-contents">Analysis of promoter leakiness and strength using flow cytometry. GFP expression in <i>Pseudomonas</i> species was analyzed by flow cytometry (<i>n</i>=7). (A) The percentage of GFP-positive cells was determined relative to the level for bacteria without a GFP plasmid. Means and standard deviations are shown. pMQ578 has <i>P<sub>xut</sub></i>, and pMQ80 has <i>P<sub>BAD</sub></i>, driving <i>gfp</i> expression. Asterisks indicate significant differences (<i>P</i>&lt;0.05) from the no-inducer group by Students <i>t</i> test. (B) Fluorescence intensity of GFP-positive cells that have been induced (inducer concentration, 10mM). Means and standard deviations are shown. Asterisks indicate significant differences (<i>P</i>&lt;0.05) between species by Students <i>t</i> test.</div>
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data-label-slug="fig-9">
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itemprop="contentUrl"
aria-describedby="figure-caption-8"
role="button"
data-image-width="750"
data-image-height="447"
data-image-alt="FIG 9"
data-pmc-id="PMC7357479"
data-figure-id="F9">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-8"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3063/7357479/59599e5a80f4/AEM.00250-20-f0009.gif"
alt="FIG 9" />
</a>
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<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 9 </p>
</strong>
<div class="figure-caption-contents"><p> XcpQ is required for the… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 9 </p>
</strong>
<div class="figure-caption-contents"><p> XcpQ is required for the inhibition of stratified corneal cell migration <i> in vitro… </i> </p></div>
</div>
<figcaption id="figure-caption-8" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 9
</strong>
<div class="figure-caption-contents">XcpQ is required for the inhibition of stratified corneal cell migration <i>in vitro</i>. Shown are representative images of cells stained with the vital stain Calcein AM and imaged by confocal microscopy. (A to C) A cell-free zone (black circle) in layers of a stratified human corneal cell line (A) closes rapidly when treated with LB medium and incubated for 24 h (B), but this migration is inhibited by filtrates from PA14 cultures grown in LB medium (C). (D) Filtrates from isogenic Δ<i>xcpQ</i> mutants were unable to impede cell migration. (E and F) <sc>d</sc>-Xylose (50mM)-induced expression of <i>gfp</i> from pMQ578 did not alter the migration phenotypes of cells exposed to PA14 or Δ<i>xcpQ</i> mutant filtrates. (G and H) Culture filtrates from the Δ<i>xcpQ</i> mutant with the wild-type <i>xcpQ</i> gene on a plasmid (pMQ644) that had been grown in LB medium with 5mM <sc>d</sc>-xylose were unable to inhibit cell migration (G), but those grown with 50mM <sc>d</sc>-xylose were complemented for the cell migration inhibition-defective phenotype (H). All cultures were grown to stationary phase and were adjusted to an OD<sub>600</sub> of 2 with fresh LB, and bacteria were removed by centrifugation and filtration.</div>
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<div id="related-links" class="related-links">
<h2 class="title">
Related information
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<ul class="related-links-list"><li><a title="Link to related Genes" href="https://www.ncbi.nlm.nih.gov/gene?linkname=pubmed_gene&amp;from_uid=32414795"
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Gene
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