Difference between revisions of "Part:BBa K1742001"

 
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<partinfo>BBa_K1742001 short</partinfo>
 
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Diarrheal diseases caused by Vibrio cholera and enterotoxigenic Escherichia coli (ETEC) are worldwide health problems that might be prevented with vaccines based on edible plants expressing the B subunit from either the cholera toxin (CTB) or the E. coli heat labile toxin (LTB). The E. coli heat-labile enterotoxin (LT) is the major ETEC pathogenic factor. LT subunit B (LTB) is of special interest because its immunogenicity, adjuvant activity and lack of toxicity make it a viable antigen to vaccinate against ETEC diarrhea.
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The heat-labile enterotoxin beta subunit is an important pathogenic factor derived from the enterotoxigenic ''Escherichia coli'' and other pathogens. It causes diarrheal diseases responsible for more than 650 million cases, resulting in almost one million deaths every year [1].
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BIOLOGY AND USAGE
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The LTB gene was domesticated and standardized as a GoldenBraid part and cloned into the pUPD2 entry vector. The CDS was subsequently assembled with the constitutive promoter P35S from Cauliflower Mosaic Virus, and its terminator (T35S). The transcriptional unit was then introduced into ''Nicotiana benthamiana'' by Agrobacterium-mediated transformation and the expression levels of LTB was detected with a Western Blot analysis employing an anti-6xHystidine antibody.
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https://static.igem.org/mediawiki/2015/thumb/5/52/BBa_K1742001_LTBexpression.png/524px-BBa_K1742001_LTBexpression.png
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<small><p><b>Figure 1. Immunoblot assay of the expression levels of LTB protein in ''N. benthamiana'' leaves.</b></p></small>
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'''References'''<br>
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1. Kang TJ., Han SC., Jank MO., Kang KH., Jang YS., Yang MS. (2004). Enhanced expression of beta subunit of Escherichia coli heat-labile enterotoxin in Tobacco by optimization of coding sequence. Applied Biochem. and Biotech. 117: 175-187<br>
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2. Mendoza SR., Soria RL., Revilla RL., Fierros LM., Alpuche AG. (2008). Ingestion of transgenic carrots expressing the Escherichia coli heat-labile enterotoxin beta subunit protects mice against cholera toxin challenge<br>
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</small>
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Revision as of 23:28, 16 September 2015

Entertoxin LTB


The heat-labile enterotoxin beta subunit is an important pathogenic factor derived from the enterotoxigenic Escherichia coli and other pathogens. It causes diarrheal diseases responsible for more than 650 million cases, resulting in almost one million deaths every year [1]. 

BIOLOGY AND USAGE

The LTB gene was domesticated and standardized as a GoldenBraid part and cloned into the pUPD2 entry vector. The CDS was subsequently assembled with the constitutive promoter P35S from Cauliflower Mosaic Virus, and its terminator (T35S). The transcriptional unit was then introduced into Nicotiana benthamiana by Agrobacterium-mediated transformation and the expression levels of LTB was detected with a Western Blot analysis employing an anti-6xHystidine antibody.

524px-BBa_K1742001_LTBexpression.png

Figure 1. Immunoblot assay of the expression levels of LTB protein in N. benthamiana leaves.

References
1. Kang TJ., Han SC., Jank MO., Kang KH., Jang YS., Yang MS. (2004). Enhanced expression of beta subunit of Escherichia coli heat-labile enterotoxin in Tobacco by optimization of coding sequence. Applied Biochem. and Biotech. 117: 175-187
2. Mendoza SR., Soria RL., Revilla RL., Fierros LM., Alpuche AG. (2008). Ingestion of transgenic carrots expressing the Escherichia coli heat-labile enterotoxin beta subunit protects mice against cholera toxin challenge


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]