Expression of Escherichia coli Heat-Labile Enterotoxin B Subunit in Centella (Centella asiatica (L.) Urban) via Biolistic Transformation.
Identifieur interne : 000150 ( Main/Exploration ); précédent : 000149; suivant : 000151Expression of Escherichia coli Heat-Labile Enterotoxin B Subunit in Centella (Centella asiatica (L.) Urban) via Biolistic Transformation.
Auteurs : Nguyen H. Loc [Viêt Nam] ; Nghiem V. Tung [Viêt Nam] ; Phan T A. Kim [Viêt Nam] ; Moon S. Yang [Corée du Sud]Source :
- Current pharmaceutical biotechnology [ 1873-4316 ] ; 2020.
Descripteurs français
- KwdFr :
- Animaux (MeSH), Biolistique (méthodes), Centella (génétique), Centella (métabolisme), Entérotoxines (composition chimique), Entérotoxines (génétique), Entérotoxines (immunologie), Entérotoxines (métabolisme), Escherichia coli (génétique), Escherichia coli (métabolisme), Expression des gènes (MeSH), Protéines Escherichia coli (génétique), Protéines Escherichia coli (immunologie), Protéines Escherichia coli (métabolisme), Température élevée (MeSH), Toxines bactériennes (génétique), Toxines bactériennes (immunologie), Toxines bactériennes (métabolisme), Triterpènes (MeSH), Végétaux génétiquement modifiés (génétique), Végétaux génétiquement modifiés (métabolisme).
- MESH :
- composition chimique : Entérotoxines.
- génétique : Centella, Entérotoxines, Escherichia coli, Protéines Escherichia coli, Toxines bactériennes, Végétaux génétiquement modifiés.
- immunologie : Entérotoxines, Protéines Escherichia coli, Toxines bactériennes.
- métabolisme : Centella, Entérotoxines, Escherichia coli, Protéines Escherichia coli, Toxines bactériennes, Végétaux génétiquement modifiés.
- méthodes : Biolistique.
- Animaux, Expression des gènes, Température élevée, Triterpènes.
English descriptors
- KwdEn :
- Animals (MeSH), Bacterial Toxins (genetics), Bacterial Toxins (immunology), Bacterial Toxins (metabolism), Biolistics (methods), Centella (genetics), Centella (metabolism), Enterotoxins (chemistry), Enterotoxins (genetics), Enterotoxins (immunology), Enterotoxins (metabolism), Escherichia coli (genetics), Escherichia coli (metabolism), Escherichia coli Proteins (genetics), Escherichia coli Proteins (immunology), Escherichia coli Proteins (metabolism), Gene Expression (MeSH), Hot Temperature (MeSH), Plants, Genetically Modified (genetics), Plants, Genetically Modified (metabolism), Triterpenes (MeSH).
- MESH :
- chemical , chemistry : Enterotoxins.
- chemical , genetics : Bacterial Toxins, Enterotoxins, Escherichia coli Proteins.
- chemical , immunology : Bacterial Toxins, Enterotoxins, Escherichia coli Proteins.
- chemical , metabolism : Bacterial Toxins, Enterotoxins, Escherichia coli Proteins.
- genetics : Centella, Escherichia coli, Plants, Genetically Modified.
- metabolism : Centella, Escherichia coli, Plants, Genetically Modified.
- methods : Biolistics.
- Animals, Gene Expression, Hot Temperature, Triterpenes.
Abstract
BACKGROUND
Heat-Labile enterotoxin B subunit (LTB) produced by Escherichia coli, a non-toxic protein subunit with potential biological properties, is a powerful mucosal and parenteral adjuvant which can induce a strong immune response against co-administered antigens.
OBJECTIVE
In the present study, LTB protein, encoded by the optimized ltb (also known synthetic ltb, s-ltb) gene in centella plant (Centella asiatica) for use as an antigen, has been discussed.
METHODS
The s-ltb gene was cloned into a plant expression vector, pMYO51, adjacent to the CaMV 35S promoter and was then introduced into centella plant by biolistic transformation. PCR amplification was conducted to determine the presence of s-ltb gene in the transgenic centella plant. The expression of s-ltb gene was analyzed by immunoblotting and quantified by ELISA. In vitro activity of LTB protein was determined by GM1-ELISA.
RESULTS
PCR amplification has found seven transgenic centella individuals. However, only five of them produced LTB protein. ELISA analysis showed that the highest amount of LTB protein detected in transgenic centella leaves was about 0.8% of the total soluble protein. GM1-ELISA assay indicated that plant LTB protein bound specifically to GM1-ganglioside, suggesting that the LTB subunits formed active pentamers.
CONCLUSION
The s-ltb gene that was successfully transformed into centella plants by the biolistic method has produced a relatively high amount of plant LTB protein in the pentameric quaternary structure that has GM1-ganglioside binding affinity, a receptor on the intestinal epithelial membrane.
DOI: 10.2174/1389201021666200226094150
PubMed: 32101119
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>Bacterial Toxins (genetics)</term>
<term>Bacterial Toxins (immunology)</term>
<term>Bacterial Toxins (metabolism)</term>
<term>Biolistics (methods)</term>
<term>Centella (genetics)</term>
<term>Centella (metabolism)</term>
<term>Enterotoxins (chemistry)</term>
<term>Enterotoxins (genetics)</term>
<term>Enterotoxins (immunology)</term>
<term>Enterotoxins (metabolism)</term>
<term>Escherichia coli (genetics)</term>
<term>Escherichia coli (metabolism)</term>
<term>Escherichia coli Proteins (genetics)</term>
<term>Escherichia coli Proteins (immunology)</term>
<term>Escherichia coli Proteins (metabolism)</term>
<term>Gene Expression (MeSH)</term>
<term>Hot Temperature (MeSH)</term>
<term>Plants, Genetically Modified (genetics)</term>
<term>Plants, Genetically Modified (metabolism)</term>
<term>Triterpenes (MeSH)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux (MeSH)</term>
<term>Biolistique (méthodes)</term>
<term>Centella (génétique)</term>
<term>Centella (métabolisme)</term>
<term>Entérotoxines (composition chimique)</term>
<term>Entérotoxines (génétique)</term>
<term>Entérotoxines (immunologie)</term>
<term>Entérotoxines (métabolisme)</term>
<term>Escherichia coli (génétique)</term>
<term>Escherichia coli (métabolisme)</term>
<term>Expression des gènes (MeSH)</term>
<term>Protéines Escherichia coli (génétique)</term>
<term>Protéines Escherichia coli (immunologie)</term>
<term>Protéines Escherichia coli (métabolisme)</term>
<term>Température élevée (MeSH)</term>
<term>Toxines bactériennes (génétique)</term>
<term>Toxines bactériennes (immunologie)</term>
<term>Toxines bactériennes (métabolisme)</term>
<term>Triterpènes (MeSH)</term>
<term>Végétaux génétiquement modifiés (génétique)</term>
<term>Végétaux génétiquement modifiés (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Enterotoxins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Bacterial Toxins</term>
<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
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<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Bacterial Toxins</term>
<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
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<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Entérotoxines</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Centella</term>
<term>Escherichia coli</term>
<term>Plants, Genetically Modified</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Centella</term>
<term>Entérotoxines</term>
<term>Escherichia coli</term>
<term>Protéines Escherichia coli</term>
<term>Toxines bactériennes</term>
<term>Végétaux génétiquement modifiés</term>
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<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Entérotoxines</term>
<term>Protéines Escherichia coli</term>
<term>Toxines bactériennes</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Centella</term>
<term>Escherichia coli</term>
<term>Plants, Genetically Modified</term>
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<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Biolistics</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Centella</term>
<term>Entérotoxines</term>
<term>Escherichia coli</term>
<term>Protéines Escherichia coli</term>
<term>Toxines bactériennes</term>
<term>Végétaux génétiquement modifiés</term>
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<term>Gene Expression</term>
<term>Hot Temperature</term>
<term>Triterpenes</term>
</keywords>
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<term>Température élevée</term>
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<front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b>
</p>
<p>Heat-Labile enterotoxin B subunit (LTB) produced by Escherichia coli, a non-toxic protein subunit with potential biological properties, is a powerful mucosal and parenteral adjuvant which can induce a strong immune response against co-administered antigens.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>OBJECTIVE</b>
</p>
<p>In the present study, LTB protein, encoded by the optimized ltb (also known synthetic ltb, s-ltb) gene in centella plant (Centella asiatica) for use as an antigen, has been discussed.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>METHODS</b>
</p>
<p>The s-ltb gene was cloned into a plant expression vector, pMYO51, adjacent to the CaMV 35S promoter and was then introduced into centella plant by biolistic transformation. PCR amplification was conducted to determine the presence of s-ltb gene in the transgenic centella plant. The expression of s-ltb gene was analyzed by immunoblotting and quantified by ELISA. In vitro activity of LTB protein was determined by GM1-ELISA.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>RESULTS</b>
</p>
<p>PCR amplification has found seven transgenic centella individuals. However, only five of them produced LTB protein. ELISA analysis showed that the highest amount of LTB protein detected in transgenic centella leaves was about 0.8% of the total soluble protein. GM1-ELISA assay indicated that plant LTB protein bound specifically to GM1-ganglioside, suggesting that the LTB subunits formed active pentamers.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>CONCLUSION</b>
</p>
<p>The s-ltb gene that was successfully transformed into centella plants by the biolistic method has produced a relatively high amount of plant LTB protein in the pentameric quaternary structure that has GM1-ganglioside binding affinity, a receptor on the intestinal epithelial membrane.</p>
</div>
</front>
</TEI>
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<DateCompleted><Year>2020</Year>
<Month>10</Month>
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<ArticleTitle>Expression of Escherichia coli Heat-Labile Enterotoxin B Subunit in Centella (Centella asiatica (L.) Urban) via Biolistic Transformation.</ArticleTitle>
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<Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Heat-Labile enterotoxin B subunit (LTB) produced by Escherichia coli, a non-toxic protein subunit with potential biological properties, is a powerful mucosal and parenteral adjuvant which can induce a strong immune response against co-administered antigens.</AbstractText>
<AbstractText Label="OBJECTIVE" NlmCategory="OBJECTIVE">In the present study, LTB protein, encoded by the optimized ltb (also known synthetic ltb, s-ltb) gene in centella plant (Centella asiatica) for use as an antigen, has been discussed.</AbstractText>
<AbstractText Label="METHODS" NlmCategory="METHODS">The s-ltb gene was cloned into a plant expression vector, pMYO51, adjacent to the CaMV 35S promoter and was then introduced into centella plant by biolistic transformation. PCR amplification was conducted to determine the presence of s-ltb gene in the transgenic centella plant. The expression of s-ltb gene was analyzed by immunoblotting and quantified by ELISA. In vitro activity of LTB protein was determined by GM1-ELISA.</AbstractText>
<AbstractText Label="RESULTS" NlmCategory="RESULTS">PCR amplification has found seven transgenic centella individuals. However, only five of them produced LTB protein. ELISA analysis showed that the highest amount of LTB protein detected in transgenic centella leaves was about 0.8% of the total soluble protein. GM1-ELISA assay indicated that plant LTB protein bound specifically to GM1-ganglioside, suggesting that the LTB subunits formed active pentamers.</AbstractText>
<AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">The s-ltb gene that was successfully transformed into centella plants by the biolistic method has produced a relatively high amount of plant LTB protein in the pentameric quaternary structure that has GM1-ganglioside binding affinity, a receptor on the intestinal epithelial membrane.</AbstractText>
<CopyrightInformation>Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Loc</LastName>
<ForeName>Nguyen H</ForeName>
<Initials>NH</Initials>
<AffiliationInfo><Affiliation>Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University, Hue, Thua Thien Hue 530000, Vietnam.</Affiliation>
</AffiliationInfo>
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<AffiliationInfo><Affiliation>Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University, Hue, Thua Thien Hue 530000, Vietnam.</Affiliation>
</AffiliationInfo>
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<ForeName>Phan T A</ForeName>
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<AffiliationInfo><Affiliation>Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University, Hue, Thua Thien Hue 530000, Vietnam.</Affiliation>
</AffiliationInfo>
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<ForeName>Moon S</ForeName>
<Initials>MS</Initials>
<AffiliationInfo><Affiliation>Division of Biological Sciences and Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Chonbuk 561-756, Korea.</Affiliation>
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<Keyword MajorTopicYN="N">heat-labile enterotoxin B subunit of E. coli (LTB)</Keyword>
<Keyword MajorTopicYN="N">plant-based vaccine</Keyword>
<Keyword MajorTopicYN="N">synthetic ltb gene (s-ltb)</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year>
<Month>09</Month>
<Day>02</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="revised"><Year>2019</Year>
<Month>11</Month>
<Day>16</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2020</Year>
<Month>01</Month>
<Day>11</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="pubmed"><Year>2020</Year>
<Month>2</Month>
<Day>27</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="medline"><Year>2020</Year>
<Month>10</Month>
<Day>21</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>2020</Year>
<Month>2</Month>
<Day>27</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
</History>
<PublicationStatus>ppublish</PublicationStatus>
<ArticleIdList><ArticleId IdType="pubmed">32101119</ArticleId>
<ArticleId IdType="pii">CPB-EPUB-104838</ArticleId>
<ArticleId IdType="doi">10.2174/1389201021666200226094150</ArticleId>
</ArticleIdList>
</PubmedData>
</pubmed>
<affiliations><list><country><li>Corée du Sud</li>
<li>Viêt Nam</li>
</country>
</list>
<tree><country name="Viêt Nam"><noRegion><name sortKey="Loc, Nguyen H" sort="Loc, Nguyen H" uniqKey="Loc N" first="Nguyen H" last="Loc">Nguyen H. Loc</name>
</noRegion>
<name sortKey="Kim, Phan T A" sort="Kim, Phan T A" uniqKey="Kim P" first="Phan T A" last="Kim">Phan T A. Kim</name>
<name sortKey="Tung, Nghiem V" sort="Tung, Nghiem V" uniqKey="Tung N" first="Nghiem V" last="Tung">Nghiem V. Tung</name>
</country>
<country name="Corée du Sud"><noRegion><name sortKey="Yang, Moon S" sort="Yang, Moon S" uniqKey="Yang M" first="Moon S" last="Yang">Moon S. Yang</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>
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