helicobacter pylori toxin gene cloning sequencing and expression（幽门螺旋杆菌毒素基因克隆测序和表达式）.docVIP

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helicobacter pylori toxin gene cloning sequencing and expression（幽门螺旋杆菌毒素基因克隆测序和表达式）.doc

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helicobacter pylori toxin gene cloning sequencing and expression（幽门螺旋杆菌毒素基因克隆测序和表达式）

Helicobacter pylori toxin gene cloning sequencing and expression Papers network: Author: production Han Feng Yan Xiaojun, SU Cheng-Zhi [Keywords] Helicobacter pylorus cells Keywords: Helicobacter pyloric bacteria cells; toxin related genes; cloning; gene expression; temperature-induced Abstract: The purpose of sub-amplified, cloned Helicobacter pylori toxin the the the gene cagA middle District cag1 cag2 using E. coli expression system. Methods PCR amplification of the cagA gene, the correct the dideoxy stop sequencing, and cloned into E. coli expression vector pBV220, controlled by PR PL promoter, was transformed into E. coli DH5 @, 42 ℃ temperature induction. Results PCR segmented amplified the CagA intermediate region gene cag1, cag2 BP 975,755 respectively, cloned into plasmid pUC19 and sequenced correctly ; build cag1 in pBV cag2 the recombinant expression vector pBVcagA1, SDSpBVcagA2 engineered bacteria induced freshmen expressing protein band, Mr.: Cag136 × 103 Cag227.9 × 103, are consistent with the expected cagA protein Mr total bacterial protein of 36% and 24%. conclusion successfully cloned segments of the the cagA middle region gene amplification, and can be highly expressed in E. coli DH5 @. Keywords: Helicobacter pylori; cytotoxin-associated gene (cagA; gene cloning; gene expression; temper-ature induce Abstract: AIM To amplify and clone the middle region of Helicobacter pylori cagA gene, and express the proteins in E.coli.METHODS After the cag1and cag2were amplified by PCR and sequencd by dideoxy methods, they were insert-ed into expression vector pBV220in which exogenous gene was controlled by PR PL promoters.The recombinant plasmids pBVcag1and pBVcag2were transformed into E.coli DH5αand induced at42 ℃ respectively to express the encoded pro ┐ teins.RESULTS The middle region of cagA was amplified and975, 755bp products were got as cag1, cag2, which were cloned into pUC19and sequenced correctly.Then, their recombination expression c