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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 : 000151

Expression 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 :

RBID : pubmed:32101119

Descripteurs français

English descriptors

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:


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Le document en format XML

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<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>
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<term>Escherichia coli Proteins (immunology)</term>
<term>Escherichia coli Proteins (metabolism)</term>
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<term>Hot Temperature (MeSH)</term>
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<term>Plants, Genetically Modified (metabolism)</term>
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<term>Animaux (MeSH)</term>
<term>Biolistique (méthodes)</term>
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<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>
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<term>Protéines Escherichia coli (immunologie)</term>
<term>Protéines Escherichia coli (métabolisme)</term>
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<term>Toxines bactériennes (immunologie)</term>
<term>Toxines bactériennes (métabolisme)</term>
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<term>Végétaux génétiquement modifiés (métabolisme)</term>
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<term>Enterotoxins</term>
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<term>Bacterial Toxins</term>
<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
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<term>Bacterial Toxins</term>
<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
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<term>Bacterial Toxins</term>
<term>Enterotoxins</term>
<term>Escherichia coli Proteins</term>
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<term>Entérotoxines</term>
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<term>Centella</term>
<term>Escherichia coli</term>
<term>Plants, Genetically Modified</term>
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<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>Protéines Escherichia coli</term>
<term>Toxines bactériennes</term>
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<term>Centella</term>
<term>Escherichia coli</term>
<term>Plants, Genetically Modified</term>
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<term>Entérotoxines</term>
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<term>Gene Expression</term>
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<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>
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<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>
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