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Construction of Sacchromyces Cerevisiae Mutants without SFA1/ADH3 Gene by Disruption

Author: SongHaoLei
Tutor: HuangJianZhong
School: Fujian Normal University
Course: Microbiology
Keywords: Saccharomyces cerevisiae Alcohol dehydrogenase ADH3 SFA1 Knockout PCR 18S rDNA
CLC: Q78
Type: Master's thesis
Year: 2007
Downloads: 266
Quote: 1
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Abstract


Yeast (Saccharomyces cerevisiae) as a traditional ethanol metabolism in bacteria and eukaryotic model organisms highly scientists favor . The use of molecular breeding tools to transform Saccharomyces cerevisiae is the development direction of ethanol producing bacteria breeding . Gene deletion mutant is the ideal material to study the metabolic regulation and signal transduction in Saccharomyces cerevisiae ethanol molecular mechanisms , application of gene knock has important theoretical value addition constructed Saccharomyces cerevisiae gene deletion mutants and thus breeding ethanol yield strains . In this paper, gene knockdown addition to the technology successfully constructed SFA1/ADH3 gene deletion Saccharomyces cerevisiae mutant two , made ??the following research : 1 , using PCR -mediated gene knockout technology , successfully built the SFA1 knockout component transformation the Saccharomyces cerevisiae YS 1 < / sub > the disposable knock in addition SFA1 two alleles , the SFA1 double gene deletion the mutant YS 1 - SFA1 . The mutant than starting the strain ethanol synthesis increased by 8.0% . 2 , resistance selection marker G418 (Kan r ) followed by knockout YS 3 yeast allele ADH3 two , successful high genetic stability ADH3 gene deletion mutants The strains of YS 3 - ADH3 . The growth behavior of the mutant under anaerobic conditions compared with the original strain showed obvious differences , is a good material to explore the ADH3 gene metabolic regulation and signaling pathway conduction mechanism . 3 , and Mode S. cerevisiae compared in Saccharomyces cerevisiae YS 3 YS 4 ADH3 and SFA1 of genes downstream regulatory sequences there are significant differences were for YS , 3 : the ADH3 upstream (97.9%), ADH3 downstream (98.8%), SFA1 upstream (98.5%), SFA1 downstream ( 99.1% ) . The YS 4 : the ADH3 upstream ( 89.1% ) the the ADH3 downstream ( 90.0% ) , SFA1 upstream (91.0%), SFA1 downstream ( 86.9%) . 4,18 S rDNA sequence and cluster analysis showed that Saccharomyces cerevisiae the YS 4 ( GenBank Accession No. : EF153844 ) , and YS 5 ( GenBank Accession No. : EF153845 ) reported in the GK3 and SCZ75578 strain has a close kinship .

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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