首页> 外文会议>International Conference on Bioinformatics and Biomedical Engineering >Disruption of PDC1 Gene to Enhance Pyruvic Acid Accumulation of Saccharomyces cerevisiae - Subtitle: PDC1 disruption for pyruvate accumulatin of S. cerevisiae
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Disruption of PDC1 Gene to Enhance Pyruvic Acid Accumulation of Saccharomyces cerevisiae - Subtitle: PDC1 disruption for pyruvate accumulatin of S. cerevisiae

机译:PDC1基因的破坏,增强酿酒酵母酿酒酵母的丙酮酸积累 - 杂志:PDC1丙二属藜亚氏菌累积蛋白的破坏

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The PDC1 gene encoding a pyruvate decarboxylase in S. cerevisiae and Kan' gene were respectively amplified by PCR using oligonucleotides which contained the restriction of NotI, EcoRI, and BglII. The two genes were digested with the same EcoRI and BgM, and linked together for inserting the Kan~r gene into the PDC1 gene while the gene disruption cassette was constructed. After transformation of the linear disruption cassette (P1K) into S. cerevisiae Y2, selected transformants were checked by PCR for correct recombination. The haploid mutant Y2-1~Δ was obtained, which could increase the yield of pyruvic acid by 176% by shaking flask cultivation as compared with the parental strain Y2.
机译:使用寡核苷酸分别通过PCR分别通过PCR分别扩增了编码肠杆菌和KAN基因中的丙酮酸脱羧酶的PDC1基因。含有寡核苷酸,其含有寡核苷酸,该寡核苷酸被含有NOTI,ECORI和BGLII的限制。用相同的EcoRI和BGM消化两种基因,并将KAN〜R基因连接到PDC1基因中,同时构建基因破坏盒。在将线性破坏盒(P1K)转化为S.Cerevisiae Y2之后,通过PCR检查选定的转化体以进行正确的重组。获得单倍体突变体Y2-1〜δ,与亲本株Y2相比,通过摇动烧瓶培养可以提高176%的丙酮酸产率。

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