Judging/Variance/UChicago

From 2009.igem.org

(Difference between revisions)
(New page: ===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 essenti...)
Line 12: Line 12:
Longtine, et al.” Additional Modules for Versatile and Economical PCR-based Gene Deletion and Modification in Saccharomyces cerevisiae.” Yeast 14, 953–961 (1998).
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.
+
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!
Thank you!

Revision as of 12:40, 17 September 2009

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