Team:TU Delft/Project/tolerance/parts


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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

TUDelft BBa solvent tolerance.png

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)

TUDelft BBa Salt tolerance.png


[1] S. Tanaka,K. Ikeda, H. Miyasaka, Enhanced Tolerance Against Salt-Stress and Freezing-Stress of Escherichia coli Cells Expressing Algal bbc1 Gene. Current Microbiology, 42:173-177 (2001)

[2] Y. Suda,T. Yoshikawa,Y. Okuda,M. Tsunemoto,S. Tanaka,K. Ikeda,H. Miyasaka,M. Watanabe,K. Sasaki,K. Harada,T. Bamba,K. Hirata, Isolation and characterization of a novel antistress gene from Chlamydomonas sp. W80 Journal of Bioscience and Bioengineering, 107(4) 352-354 (2009)

[3] Y. Hase, S. Yokoyama, A. Muto, et al. , Removal of a ribosome small subunit-dependent GTPase confers salt resistance on Escherichia coli cells RNA Society, 15:1766-1774 (2009)