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Judging/Variance/UChicago
From 2009.igem.org
Team Request
Dear iGEM headquarters:
We would like to use two news types of S. cerevisae expression vectors, pRS314 and pRS316 modified to meet biobrick standards. These are essentially empty vectors with ampicillin resistance for selection in E. coli and tryptophan and uracil (respectively) in S. cerevisiae. The plasmid maps can be found at
https://www.lablife.org/ct?f=c&a=showvecinfo&vectorid=3297 https://www.lablife.org/ct?f=c&a=showvecinfo&vectorid=3299.
We are in the process of modifying these plasmids to fit biobrick standards, including removing extra XbeI, SpeI and PstI sites.
Our second request is for a new standard of yeast expression derived from
Longtine, et al.” Additional Modules for Versatile and Economical PCR-based Gene Deletion and Modification in Saccharomyces cerevisiae.” Yeast 14, 953–961 (1998).
The modules described in the paper allow for easy deletion, replacement, modification or over-expression of a gene of interest without use of a plasmid. We have been using pFA6a-GFP(S65T)-kanMX6 and pFA6a-GFP(S56T)-His3MX6. Using PCR, we amplify our module of interest with tails on either end that correspond to the gene region we wish to exploit. To integrate these fragments into the S. cerevisiae genome we simply transform with our amplified product. We hope to submit these plasmids with the Longtine modules, as well as our two biobrick-compatible cerevisiae plasmids to the registry. Please let us know whether this will be feasible.
Thank you!
Nora Yucel University of Chicago iGEM team
