<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>1025-028X</journal-id>
<journal-title><![CDATA[Vaccimonitor]]></journal-title>
<abbrev-journal-title><![CDATA[Vaccimonitor]]></abbrev-journal-title>
<issn>1025-028X</issn>
<publisher>
<publisher-name><![CDATA[Finlay Ediciones]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S1025-028X2020000300136</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Expression in Saccharomyces boulardii of recombinant Toxin-coregulated pilus A subunit (TcpA) of Vibrio cholerae O1]]></article-title>
<article-title xml:lang="es"><![CDATA[Expresión recombinante en Saccharomyces boulardii de la subunidad A del pili corregulado con la Toxina (TcpA) de Vibrio cholerae O1]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Awad]]></surname>
<given-names><![CDATA[Suadad]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Al-Ahmer]]></surname>
<given-names><![CDATA[Saife D]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Salih]]></surname>
<given-names><![CDATA[Shahla'a M]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Ministery of Science and Technology  ]]></institution>
<addr-line><![CDATA[Baghdad ]]></addr-line>
<country>Iraq</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Institute of Genetic Engineering and Biotechnology for Post Graduate study, Baghdad University  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Iraq</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,College of Biotechnology, Al-Nahrain University, Baghdad  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Iraq</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<volume>29</volume>
<numero>3</numero>
<fpage>136</fpage>
<lpage>142</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S1025-028X2020000300136&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S1025-028X2020000300136&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S1025-028X2020000300136&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Cholera is endemic in over 50 countries with an estimated mortality of 100,000-120,000. Vaccination is considered the complementary key to prevent and control cholera; therefore, alternative vaccine preparations are needed. Toxin Co-regulated Pilus is part of the toxR virulence regulon, which is necessary for colonization in the intestinal mucosa. In order to express Vibrio cholerae TcpA protein in Saccharomyces boulardii, the expression plasmid pYES2 was constructed by inserting tcpA gene isolated from local Vibrio cholerae Eltor Inaba isolates. The new construct was transferred into Saccharomyces boulardii cells and the expression of tcpA gene was induced from the GAL1 promoter by adding galactose to the medium. The SDS-PAGE and Western blot analysis showed the presence of TcpA in yeast. These results showed that Saccharomyces boulardii is a promising host to express Vibrio cholerae toxin TcpA as the first step in attempt to produce an oral Vibrio cholerae vaccine.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN El cólera es endémico en más de 50 países. Se estima una mortalidad entre 100.000 - 120.000 debido a esta enfermedad. La vacunación se considera una medida complementaria para prevenir y controlar el cólera, por lo tanto, se necesitan preparaciones vacunales alternativas a las existentes. El Pili corregulado con la toxina, es parte del regulón de virulencia toxR, y es necesario para la colonización en la mucosa intestinal. Para expresar la proteína tcpA de Vibrio cholerae en Saccharomyces boulardii, se construyó el plásmido de expresión pYES2 insertando el gen tcpA obtenido a partir de aislamientos locales de Vibrio cholerae El Tor Inaba. La nueva construcción se transfirió a las células de Saccharomyces boulardii y se indujo la expresión del gen tcpA a partir del promotor GAL1 mediante la adición de galactosa al medio. El análisis mediante SDS-PAGE y Western blot demostró la presencia de TcpA en levaduras. Los resultados demostraron que Saccharomyces boulardii es un hospedero prometedor para expresar el gen tcpA de Vibrio cholerae como el primer paso en el intento de producir una vacuna oral contra Vibrio cholerae.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[Saccharomyces boulardii]]></kwd>
<kwd lng="en"><![CDATA[Vibrio cholerae O1]]></kwd>
<kwd lng="en"><![CDATA[cholera]]></kwd>
<kwd lng="en"><![CDATA[cloning]]></kwd>
<kwd lng="en"><![CDATA[DNA]]></kwd>
<kwd lng="en"><![CDATA[Escherichia coli]]></kwd>
<kwd lng="es"><![CDATA[Saccharomyces boulardii]]></kwd>
<kwd lng="es"><![CDATA[Vibrio cholera O1]]></kwd>
<kwd lng="es"><![CDATA[cólera]]></kwd>
<kwd lng="es"><![CDATA[clonaje]]></kwd>
<kwd lng="es"><![CDATA[ADN]]></kwd>
<kwd lng="es"><![CDATA[Escherichia coli]]></kwd>
</kwd-group>
</article-meta>
</front><back>
<ref-list>
<ref id="B1">
<label>1</label><nlm-citation citation-type="book">
<collab>World Health Organization</collab>
<source><![CDATA[Newsroom Fact sheets Detail Cholera]]></source>
<year>2019</year>
<publisher-loc><![CDATA[Ginebra ]]></publisher-loc>
<publisher-name><![CDATA[WHO]]></publisher-name>
</nlm-citation>
</ref>
<ref id="B2">
<label>2</label><nlm-citation citation-type="">
<collab>Health cluster Iraq</collab>
<source><![CDATA[Wash cluster. Iraq Health and WASH Cluster Acute Diarrheal Disease (including Cholera) Preparedness and Response Plan]]></source>
<year>2018</year>
</nlm-citation>
</ref>
<ref id="B3">
<label>3</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Benenson]]></surname>
<given-names><![CDATA[AS]]></given-names>
</name>
<name>
<surname><![CDATA[Joseph]]></surname>
<given-names><![CDATA[PR]]></given-names>
</name>
<name>
<surname><![CDATA[Oseasohn]]></surname>
<given-names><![CDATA[RO]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Cholera vaccine field trials in East Pakistan 1. Reaction and antigenicity studies]]></article-title>
<source><![CDATA[Bull World Health Organ]]></source>
<year>1968</year>
<volume>38</volume>
<numero>3</numero>
<issue>3</issue>
<page-range>347-57</page-range></nlm-citation>
</ref>
<ref id="B4">
<label>4</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Bishop]]></surname>
<given-names><![CDATA[AL]]></given-names>
</name>
<name>
<surname><![CDATA[Camilli]]></surname>
<given-names><![CDATA[A]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Vibrio cholerae lessons for mucosal vaccine design]]></article-title>
<source><![CDATA[Expert Rev Vaccine]]></source>
<year>2011</year>
<volume>10</volume>
<numero>1</numero>
<issue>1</issue>
<page-range>79-94</page-range></nlm-citation>
</ref>
<ref id="B5">
<label>5</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Kelesidis]]></surname>
<given-names><![CDATA[T]]></given-names>
</name>
<name>
<surname><![CDATA[Pothoulakis]]></surname>
<given-names><![CDATA[C]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Efficacy and safety of the probiotic Saccharomyces boulardii for the prevention and therapy of gastrointestinal disorders]]></article-title>
<source><![CDATA[Ther Adv Gastroenterol]]></source>
<year>2012</year>
<volume>5</volume>
<numero>2</numero>
<issue>2</issue>
<page-range>111-25</page-range></nlm-citation>
</ref>
<ref id="B6">
<label>6</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Sazawal]]></surname>
<given-names><![CDATA[S]]></given-names>
</name>
<name>
<surname><![CDATA[Hiremath]]></surname>
<given-names><![CDATA[G]]></given-names>
</name>
<name>
<surname><![CDATA[Dhingra]]></surname>
<given-names><![CDATA[U]]></given-names>
</name>
<name>
<surname><![CDATA[Malik]]></surname>
<given-names><![CDATA[P]]></given-names>
</name>
<name>
<surname><![CDATA[Deb]]></surname>
<given-names><![CDATA[S]]></given-names>
</name>
<name>
<surname><![CDATA[Black]]></surname>
<given-names><![CDATA[RE]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Efficacy of probiotics in prevention of acute diarrhoea a meta-analysis of masked, randomised, placebo-controlled trials]]></article-title>
<source><![CDATA[Lancet Infect Dis]]></source>
<year>2006</year>
<volume>6</volume>
<page-range>374-82</page-range></nlm-citation>
</ref>
<ref id="B7">
<label>7</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Douradinha]]></surname>
<given-names><![CDATA[B]]></given-names>
</name>
<name>
<surname><![CDATA[Reis]]></surname>
<given-names><![CDATA[VC]]></given-names>
</name>
<name>
<surname><![CDATA[Rogers]]></surname>
<given-names><![CDATA[MB]]></given-names>
</name>
<name>
<surname><![CDATA[Torres]]></surname>
<given-names><![CDATA[FA]]></given-names>
</name>
<name>
<surname><![CDATA[Evans]]></surname>
<given-names><![CDATA[JD]]></given-names>
</name>
<name>
<surname><![CDATA[Marques Jr]]></surname>
<given-names><![CDATA[ET]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Novel insights in genetic transformation of the probiotic yeast Saccharomyces boulardii]]></article-title>
<source><![CDATA[Bioengineered]]></source>
<year>2014</year>
<volume>5</volume>
<numero>1</numero>
<issue>1</issue>
<page-range>21-9</page-range></nlm-citation>
</ref>
<ref id="B8">
<label>8</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Vohra]]></surname>
<given-names><![CDATA[A]]></given-names>
</name>
<name>
<surname><![CDATA[Satyanarayana]]></surname>
<given-names><![CDATA[T]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Probiotic yeasts]]></article-title>
<person-group person-group-type="editor">
<name>
<surname><![CDATA[Satyanarayana]]></surname>
<given-names><![CDATA[T]]></given-names>
</name>
</person-group>
<source><![CDATA[Microorganisms in sustainable agriculture and biotechnology]]></source>
<year>2012</year>
<page-range>411-33</page-range><publisher-loc><![CDATA[Dordrecht ]]></publisher-loc>
<publisher-name><![CDATA[Springer]]></publisher-name>
</nlm-citation>
</ref>
<ref id="B9">
<label>9</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Liu]]></surname>
<given-names><![CDATA[JJ]]></given-names>
</name>
<name>
<surname><![CDATA[Kong]]></surname>
<given-names><![CDATA[II]]></given-names>
</name>
<name>
<surname><![CDATA[Zhang]]></surname>
<given-names><![CDATA[GC]]></given-names>
</name>
<name>
<surname><![CDATA[Jayakody]]></surname>
<given-names><![CDATA[LN]]></given-names>
</name>
<name>
<surname><![CDATA[Kim]]></surname>
<given-names><![CDATA[H]]></given-names>
</name>
<name>
<surname><![CDATA[Xia]]></surname>
<given-names><![CDATA[PF]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Metabolic engineering of probiotic Saccharomyces boulardii]]></article-title>
<source><![CDATA[Appl Environ Microbiol]]></source>
<year>2016</year>
<volume>82</volume>
<page-range>2280-7</page-range></nlm-citation>
</ref>
<ref id="B10">
<label>10</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Khatri]]></surname>
<given-names><![CDATA[I]]></given-names>
</name>
<name>
<surname><![CDATA[Tomar]]></surname>
<given-names><![CDATA[R]]></given-names>
</name>
<name>
<surname><![CDATA[Ganesan]]></surname>
<given-names><![CDATA[K]]></given-names>
</name>
<name>
<surname><![CDATA[Prasad]]></surname>
<given-names><![CDATA[GS]]></given-names>
</name>
<name>
<surname><![CDATA[Subramanian]]></surname>
<given-names><![CDATA[S]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Complete genome sequence and comparative genomics of the probiotic yeast Saccharomyces boulardii]]></article-title>
<source><![CDATA[Sci Rep]]></source>
<year>2017</year>
<volume>7</volume>
<numero>1</numero>
<issue>1</issue>
<page-range>371</page-range></nlm-citation>
</ref>
<ref id="B11">
<label>11</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Krebs]]></surname>
<given-names><![CDATA[SJ]]></given-names>
</name>
<name>
<surname><![CDATA[Taylor]]></surname>
<given-names><![CDATA[RK]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Protection and attachment of Vibrio cholerae mediated by the toxin-coregulated pilus in the infant mouse model]]></article-title>
<source><![CDATA[J Bacteriol]]></source>
<year>2011</year>
<volume>193</volume>
<numero>19</numero>
<issue>19</issue>
<page-range>5260-70</page-range></nlm-citation>
</ref>
<ref id="B12">
<label>12</label><nlm-citation citation-type="book">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Forbes]]></surname>
<given-names><![CDATA[BA]]></given-names>
</name>
<name>
<surname><![CDATA[Sahm]]></surname>
<given-names><![CDATA[DF]]></given-names>
</name>
<name>
<surname><![CDATA[Weissfeld]]></surname>
<given-names><![CDATA[AS]]></given-names>
</name>
</person-group>
<source><![CDATA[Study guide for Bailey &amp; Scott&amp;apos;s diagnostic microbiology]]></source>
<year>2007</year>
<publisher-loc><![CDATA[Saint Louis ]]></publisher-loc>
<publisher-name><![CDATA[Elsevier Mosby]]></publisher-name>
</nlm-citation>
</ref>
<ref id="B13">
<label>13</label><nlm-citation citation-type="book">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Kaysner]]></surname>
<given-names><![CDATA[CA]]></given-names>
</name>
<name>
<surname><![CDATA[DePaola]]></surname>
<given-names><![CDATA[A]]></given-names>
</name>
<name>
<surname><![CDATA[Jones]]></surname>
<given-names><![CDATA[J]]></given-names>
</name>
</person-group>
<source><![CDATA[Bacteriological Analytical Manual (BAM) Chapter 9: Vibrio]]></source>
<year>2004</year>
<publisher-name><![CDATA[FDA]]></publisher-name>
</nlm-citation>
</ref>
<ref id="B14">
<label>14</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Goforth]]></surname>
<given-names><![CDATA[JB]]></given-names>
</name>
<name>
<surname><![CDATA[Walter]]></surname>
<given-names><![CDATA[NE]]></given-names>
</name>
<name>
<surname><![CDATA[Karatan]]></surname>
<given-names><![CDATA[E]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Effects of polyamines on Vibrio cholerae virulence properties]]></article-title>
<source><![CDATA[PLoS One]]></source>
<year>2013</year>
<volume>8</volume>
<numero>4</numero>
<issue>4</issue>
</nlm-citation>
</ref>
<ref id="B15">
<label>15</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Hudson]]></surname>
<given-names><![CDATA[LE]]></given-names>
</name>
<name>
<surname><![CDATA[Fasken]]></surname>
<given-names><![CDATA[MB]]></given-names>
</name>
<name>
<surname><![CDATA[McDermott]]></surname>
<given-names><![CDATA[CD]]></given-names>
</name>
<name>
<surname><![CDATA[McBride]]></surname>
<given-names><![CDATA[SM]]></given-names>
</name>
<name>
<surname><![CDATA[Kuiper]]></surname>
<given-names><![CDATA[EG]]></given-names>
</name>
<name>
<surname><![CDATA[Guiliano]]></surname>
<given-names><![CDATA[DB]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Functional heterologous protein expression by genetically engineered probiotic yeast Saccharomyces boulardii]]></article-title>
<source><![CDATA[PloS One]]></source>
<year>2014</year>
<volume>9</volume>
<numero>11</numero>
<issue>11</issue>
</nlm-citation>
</ref>
<ref id="B16">
<label>16</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Benatuil]]></surname>
<given-names><![CDATA[L]]></given-names>
</name>
<name>
<surname><![CDATA[Perez]]></surname>
<given-names><![CDATA[JM]]></given-names>
</name>
<name>
<surname><![CDATA[Belk]]></surname>
<given-names><![CDATA[J]]></given-names>
</name>
<name>
<surname><![CDATA[Hsieh]]></surname>
<given-names><![CDATA[CM]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[An improved yeast transformation method for the generation of very large human antibody libraries]]></article-title>
<source><![CDATA[Protein Eng Des Sel]]></source>
<year>2010</year>
<volume>23</volume>
<numero>4</numero>
<issue>4</issue>
<page-range>155-9</page-range></nlm-citation>
</ref>
<ref id="B17">
<label>17</label><nlm-citation citation-type="book">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Sambrook]]></surname>
<given-names><![CDATA[J]]></given-names>
</name>
<name>
<surname><![CDATA[Russell]]></surname>
<given-names><![CDATA[DW]]></given-names>
</name>
</person-group>
<source><![CDATA[Molecular cloning: A laboratory manual]]></source>
<year>2001</year>
<publisher-loc><![CDATA[New York ]]></publisher-loc>
<publisher-name><![CDATA[Cold Spring Harbor Laboratory Press]]></publisher-name>
</nlm-citation>
</ref>
<ref id="B18">
<label>18</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Mahmood]]></surname>
<given-names><![CDATA[T]]></given-names>
</name>
<name>
<surname><![CDATA[Yang]]></surname>
<given-names><![CDATA[PC]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Western blot technique, theory, and trouble shooting]]></article-title>
<source><![CDATA[N Am J Med Sci]]></source>
<year>2012</year>
<volume>4</volume>
<page-range>429-34</page-range></nlm-citation>
</ref>
<ref id="B19">
<label>19</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Lim]]></surname>
<given-names><![CDATA[JG]]></given-names>
</name>
<name>
<surname><![CDATA[Jin]]></surname>
<given-names><![CDATA[HS]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Heterologous expression of cholera toxin B subunit in Saccharomyces cerevisiae]]></article-title>
<source><![CDATA[Biotechnol Bioprocess E]]></source>
<year>2008</year>
<volume>13</volume>
<page-range>598-605</page-range></nlm-citation>
</ref>
<ref id="B20">
<label>20</label><nlm-citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname><![CDATA[Bagherpour]]></surname>
<given-names><![CDATA[G]]></given-names>
</name>
<name>
<surname><![CDATA[Ghasemi]]></surname>
<given-names><![CDATA[H]]></given-names>
</name>
<name>
<surname><![CDATA[Zand]]></surname>
<given-names><![CDATA[B]]></given-names>
</name>
<name>
<surname><![CDATA[Zarei]]></surname>
<given-names><![CDATA[N]]></given-names>
</name>
<name>
<surname><![CDATA[Roohvand]]></surname>
<given-names><![CDATA[F]]></given-names>
</name>
<name>
<surname><![CDATA[Ardakani]]></surname>
<given-names><![CDATA[EM]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Oral administration of recombinant Saccharomyces boulardii expressing ovalbumin-CPE fusion protein induces antibody response in mice]]></article-title>
<source><![CDATA[Front microbiol]]></source>
<year>2018</year>
<volume>9</volume>
<page-range>723</page-range></nlm-citation>
</ref>
</ref-list>
</back>
</article>
