Homework Search for 1 paper that has used gene knockout technique to study yeast
ID: 200733 • Letter: H
Question
Homework Search for 1 paper that has used gene knockout technique to study yeast gene function. Outline the method, gene function and conclusion. . I wish to obtain a yeast knockout strain, FPS1 knockout strain, please search for the available resource and list the 1) website 2) Catalog number and 3) the selective marker they used Design step-wise approach to knockout a gene in yeast. Choose any gene that you are interested (not a lethal gene though). Include the gene sequence, regions that you wish to remove, primers that you are using and optional if you choose to do the homework from last class).Explanation / Answer
Paper: Akada et al. (2006).PCR-mediated seamless gene deletion and marker recycling in Saccharomyces cerevisiae. Retrieved from https://www.google.co.in/url?sa=t&source=web&rct=j&url=http://onlinelibrary.wiley.com/doi/10.1002/yea.1365/pdf&ved=2ahUKEwjix5qf38HZAhWHPY8KHV8PDekQFjACegQICBAB&usg=AOvVaw2oK0R8tn3o-qOWzoeegTCd
Target gene: His3
Gene function: Histidine biosynthesis
Method: URA3 was used as a marker. His3 40 bases sequence was obtained and tagged with URA3 on both sides of this sequence. The gene was targetted by a 5' homologous sequence carried out by fusion PCR. The fragments were transfected into the yeast strain BY4740 and transformed colonies were identified by colony PCR. The transformed colonies had deleted His3 region and showed no expression of the reporter gene URA3. Authors conclude that PCR amplification method is useful in generating target gene deletions using primers that contain the target gene sequence.
(only first question answered)
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