Team:TU Delft/Project/tolerance/parts

From 2010.igem.org

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(New page: '''== The Biobricks ==''' Genes for solvent and salt tolerance were found during our literature review: BBC1, prefoldin alpha and beta. The original sequences of these genes differ in c...)
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==The Biobricks==
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'''== The Biobricks =='''
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Genes for solvent and salt tolerance were found during our literature review: BBC1, prefoldin alpha and beta. The original sequences of these genes differ in composition from the E. coli sequences; furthermore they contain undesired restriction sites. We decided to enhance our sequences for a successful expression on E. coli, for that purpose we used jcat website tool.  
Genes for solvent and salt tolerance were found during our literature review: BBC1, prefoldin alpha and beta. The original sequences of these genes differ in composition from the E. coli sequences; furthermore they contain undesired restriction sites. We decided to enhance our sequences for a successful expression on E. coli, for that purpose we used jcat website tool.  
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Once we had the enhanced sequence, we added the standard Biobric prefix and suffix. Finally, we sticked the promoter-RBS sequences that best suit for our CDS.  
Once we had the enhanced sequence, we added the standard Biobric prefix and suffix. Finally, we sticked the promoter-RBS sequences that best suit for our CDS.  
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Solvent tolerance  
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'''Solvent tolerance'''
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For solvent tolerance the following construct was made:
For solvent tolerance the following construct was made:
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J23100- phPFDα
J23100- phPFDα
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J23100 phPFDβ
J23100 phPFDβ
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J23100- phPFDα – J23100 phPFDβ  
J23100- phPFDα – J23100 phPFDβ  
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'''Salt Tolerance'''
For salt tolerance the following constructs were made:
For salt tolerance the following constructs were made:
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J23100- bbc1-B0015 (Low expression)
J23100- bbc1-B0015 (Low expression)
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J23101-bbc1-B0015 (Low to moderate expression
J23101-bbc1-B0015 (Low to moderate expression
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J23119-B0034-bbc1-B0015 (High expression)
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J23119-B0034-bbc1-B0015         (High expression)

Revision as of 10:52, 12 September 2010

The Biobricks

Genes for solvent and salt tolerance were found during our literature review: BBC1, prefoldin alpha and beta. The original sequences of these genes differ in composition from the E. coli sequences; furthermore they contain undesired restriction sites. We decided to enhance our sequences for a successful expression on E. coli, for that purpose we used jcat website tool.

Once we had the enhanced sequence, we added the standard Biobric prefix and suffix. Finally, we sticked the promoter-RBS sequences that best suit for our CDS.


Solvent tolerance

For solvent tolerance the following construct was made:

J23100- phPFDα

J23100 phPFDβ

J23100- phPFDα – J23100 phPFDβ

Salt Tolerance

For salt tolerance the following constructs were made:

J23100- bbc1-B0015 (Low expression)

J23101-bbc1-B0015 (Low to moderate expression

J23119-B0034-bbc1-B0015 (High expression)