文档介绍：Disruption of PDC1 Gene to Enhance Pyruvic Acid
Accumulation of haromyces cerevisiae
Subtitle: PDC1 disruption for pyruvate accumulatin of S. cerevisiae
WANG De-pei1,2 (
), DING Xin1,2, DENG Xu-heng1,3, ZHAO Qin1,3, GAO Qiang1, 2, GAO Nian-fa1,2 (
)
1. School of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, P. R. China;
2. Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin 300457, P. R. China;
3. Tianjin Key Laboratory of Industrial Microbiology, Tianjin 300457, P. R. China
E-mails: ******@tust.; gaonianfa@
Abstract—The PDC1 gene encoding a pyruvate decarboxylase pPIC- were gifts from Nankai University, pMD19-T
in S. cerevisiae and Kanr gene were respectively amplified by PCR Simple was bought from TAKARA Co., Ltd. The three
using oligonucleotides which contained the restriction of NotI, plasmids all exhibited ampicillin resistance in E. coli.
EcoRI, and BglII. The two genes were digested with the same
EcoRI and BglII, and linked together for inserting the Kanr gene B. Media and Culture Conditions
into the PDC1 gene while the gene disruption cassette was S. cerevisiae Y2 strain was grown at 30°C on YPD
constructed. After transformation of the linear disruption cassette medium (1% yeast extract, 2% peptone, 2% glucose) for
(P1K) into S. cerevisiae Y2, selected transformants were