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[目的 ]在大肠杆菌中克隆及高效表达猪囊尾蚴抗原cC1。 [方法 ]将cC1cDNA片段以BamHI和PstI克隆到pGEM 3Z载体 ,改换限制性内切酶位点成BamHI和PstI,加上人工接头PstI BamHI XhoI后 ,克隆到pGEX 5T ,构建重组原核表达载体pGE X5T cC1。 [结果 ]培养 3h、IPTG诱导 6h ,cC1表达量最高 ,表达量占细菌总量的 5 7% ,Westernblotting结果表明猪囊尾蚴抗原cC1蛋白与囊尾蚴病猪血清有特异性的结合条带。 [结论 ]猪囊尾蚴抗原cC1在大肠杆菌中获得高效表达。
[Objective] The research aimed to clone and express cysticercus cellulosae antigen cC1 in Escherichia coli. [Method] The cCl cDNA fragment was cloned into pGEM 3Z vector with BamHI and PstI, the restriction endonuclease site was changed into BamHI and PstI, and then the artificial linker PstI BamHI XhoI was cloned into pGEX 5T to construct recombinant prokaryotic expression vector pGE X5T cC1. [Result] After induced by IPTG for 6h, the expression level of cC1 was the highest, accounting for 57.7% of the total bacteria. The results of Western blotting showed that there was a specific binding band between cC1 antigen of cysticercus cellulosae and pig serum of cysticercosis. [Conclusion] The cysticercus cellulosa antigen cC1 was highly expressed in E. coli.