Xiangling Chen, Hanying Yuan, Wei He, Xianghua Hu, Hong Lu, Yuyang Li
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
Science in China. Series C, Life sciences / Chinese Academy of Sciences 2005 AugBased on a previously used plasmid pHC11, a new plasmid pHC11R was constructed. Cutting plasmid pHC11R with proper restriction enzymes, the resulting larger DNA fragment pHC11R' was co-transformed with a PCR amplified expression cassette of human IFNalpha2b into yeast. By means of the homologous sequences at both ends of two DNA fragments, a novel expression plasmid pHC11R-IFNalpha2b was formed via homologous recombination in the yeast. Compared with pHC11-IFNalpha2b, the expression plasmid pHC11R-IFNalpha2b was smaller in size and in absence of antibiotic resistant gene. The stability and copy number of pHC11R-IFNalpha2b were greatly increased and the expression level of heterologous protein was improved. As the derivatives of pHC11R, a series of recombination expression vectors pHRs containing different combination of expression elements were developed. This led to a rapid and powerful method for cloning and expressing of different genes in yeast.
Xiangling Chen, Hanying Yuan, Wei He, Xianghua Hu, Hong Lu, Yuyang Li. Construction of a novel kind of expression plasmid by homologous recombination in Saccharomyces cerevisiae. Science in China. Series C, Life sciences / Chinese Academy of Sciences. 2005 Aug;48(4):330-6
PMID: 16248426
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