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2013年50 No.2
Cloning, expression of cytochrome P450 from Locusta migratoria manilensis
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Key Words：Locusta migratoria manilensis, cytochrome P450, gene clone, expression
Abstract：To clone the gene of cytochrome P450 (CYP450) from Locusta migratoria manilensis (Meyen)，produce its recombinant
protein.The cDNA of CYP450 was cloned using specific primers from the total RNA of L. m. manilensis. The cloned gene was
inserted into pMD18T vector and digested by BamH I and Hind III. The cDNA was sequenced and subcloned into pET28a
expression vector. The cloned CYP450 cDNA gene was expressed in Escherichia coli Rosetta by IPTG induction. Result showed
that the cloned cDNA ORF sequence contained 1 551 bp and encoded 516 amino acids. Its sequence homology with the published
one (Accession no. HM153426) was 99% at nucleotide level. The CYP450 was highly expressed in E. coli Rosetta as a unsoluble
protein mainly with the molecular weight of about Mr 53 000 under induction of IPTG.