Building method of pichia pastoris gene engineering strain for producing lipase
A technology of genetically engineered strains and construction methods, applied in the field of constructing genetically engineered strains of Pichia pastoris, can solve the problems of high price and high cost of biodiesel
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[0025]1. The construction method of the Pichia genetically engineered strain producing lipase is characterized in that it is carried out in the following steps: one, the lipase gene is derived from the genome of Candida sp.99-125; the Candida sp.99-125 genome is extracted by the extraction kit Candida sp.99-125 genome; 2. PCR amplification of Lip14 gene: PCR amplification was performed using the Candida sp.99-125 genome extracted in step 1 as a template, the product was detected by electrophoresis, and the target gene band Purify and recover the target gene Lip14 by gel recovery kit; the forward primer for PCR amplification is 5'-GGAATTCGTTCTATTCTACAGTCATTGGAGAATGGT-3', and the reverse primer is 5'-GCTCTAGAAAAAAGATGGTGGAACCGTTGG-3' 3. Recombinant expression vector pPICZα-Lip14 Construction: The Lip14 gene purified in step 2 was digested with EcoR I and Xba I, and then purified and recovered by a PCR recovery kit. The expression vector pPICZαA was double digested with EcoR I and...
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