Application of Droplet Digital PCR in Screening of Genetically Modified Saccharomyces cerevisiae for Multicopy Expression of Xylanase

doi:10.7506/spkx1002-6630-201810028

Abstract

Abstract: In order to obtain a high yield of xylanase from genetically engineered Saccharomyces cerevisiae, the S. cerevisiae expression vector for the multicopy expression of the xylanase gene was constructed by rDNA integration method. Then, the copy number of transformants was detected by droplet digital PCR, and the relationship between the copy number of the xylanase gene and the enzyme activity was analyzed. The results indicated that 10 strains of S. cerevisiae were obtained by rDNA integration method. Their enzymatic activity was measured. The xylanase production capacity was increased to the maximum of 308 U/mL with copy number up to 9, but decreased when the copy number was more than 9.

Key words: droplet digital PCR, multicopy transformant, Saccharomyces cerevisiae, xylanase, rDNA integration method

CLC Number:

Q936

LAN Xue, ZHANG Sitong, LI Zhe, CHANG Hao, SUN Yang, WANG Gang, CHEN Huan, WANG Chunfeng, CHEN Guang. Application of Droplet Digital PCR in Screening of Genetically Modified Saccharomyces cerevisiae for Multicopy Expression of Xylanase[J]. FOOD SCIENCE, 2018, 39(10): 179-184.

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Application of Droplet Digital PCR in Screening of Genetically Modified Saccharomyces cerevisiae for Multicopy Expression of Xylanase

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