Lactic dehydrogenase humanization saccharomyces cerevisiae and construction method thereof
2015
The invention discloses lactic dehydrogenase humanization saccharomyces cerevisiae and a construction method thereof, and belongs to the technical field of microorganism and technical field of molecular biology. Genes PDC1, PDC5 and PDC6 of a saccharomyces cerevisiae code PDC are knocked out by adopting a gene homologous recombination method, LDH derived from the human being is expressed in the saccharomyces cerevisiae, the transmission of an electron chain is blocked by utilizing antimycin A, so that the humanization lactic acid fermentation substitutes the alcoholic fermentation to maintain glycolysis. By adopting the lactic dehydrogenase humanization saccharomyces cerevisiae, the heterologous expression of lactic dehydrogenase is realized. The lactic dehydrogenase humanization saccharomyces cerevisiae can express the lactic dehydrogenase of the human being, LDHA or LDHC can be expressed in three defected strains pdc1 delta, pdc6 delta and pdc5 delta of the saccharomyces cerevisiae, and growth can be realized and the lactic acid can be detected by adding the antimycin A. The lactic dehydrogenase humanization saccharomyces cerevisiae can be used as a potential tool for screening the anticancer drugs and adjusting genes associated to the activity of the lactic dehydrogenase.
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