Chitinase Gene Overexpression of Chaetomium cupreum in Pichia pastoris by Multi-site Mutations
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    Abstract:

    By using gene SOEing PCR, Arg (CGC) in the expression fragment of chitinase gene (chi58) from Chaetomium cupreum was mutated synonymously to Arg (AGA), which is a bias code of Pichia pastoris yeast. The expression plasmid pPIC9K-chi58A was constructed and transformed into GS115 strain through electroporation. The chitinase activity reached 101.71 U/mL±3.33 U/mL after induced with 120 h, which was the 3 fold of the original strain (31.83 U/mL±4.85 U/mL). Additionally, recombinant yeast showed good genetic stability after 10 cycle. SDS-PAGE analysis suggested that the weight of protein was 58 kD.

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WANG Yan-Jun, YANG Qian. Chitinase Gene Overexpression of Chaetomium cupreum in Pichia pastoris by Multi-site Mutations[J]. Microbiology China, 2008, 35(10): 1544-1549

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