人LOX-1基因毕赤酵母表达载体的构建与表达研究

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Abstract Objective To construct LOX-1 eukaryotic expression vector and screen high expression strain,lay the foundation for obtaining active recombinant LOX-1 protein in vitro.M ethods LOX-1 gene was amplified by PCR with the plasmid containing human LOX-1 gene as a template.After TA clone,LOX-1 gene was subcloned into eukaryotic expression vector pPICZαA to obtain pPICZαA-LOX-1 recombinant plasmid.After double digestion identification,the positive recombinant plasmid was electro transformed into Pichia pastoris X-33.Situ double membrane hybridization method was used to rapidly screen positive highly expressed strains.Finally,Western blotwas used to test its expression. Results LOX-1 recombinant expressed vector was constructed and highly expressed strain was screened out successfully.Western blot results showed that the LOX-1 recombinant protein was expressed in the cell culture supernatant after induction by methanol.Conclusion LOX-1 gene is expressed in Pichia pastoris cell supernatant.This study laies a foundation for obtaining active recombinant LOX-1 protein in vitro and studying its role in atherosclerosis.

Key words： LOX-1 Vector construction Pichia pastoris Atherosclerosis

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	Articles by authors
	LIU Jing
	YU Tian-tian
	CHEN Yu-hua
	LI Jing-da
	TAO Ya-ni
	LI Ming-ying
	WANG Xiao
	CHEN Yong
	CHI Yan

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LIU Jing,YU Tian-tian,CHEN Yu-hua, et al. Study of the establishment of human LOX-1 gene Pichia expression vector and its expression[J]. 中国当代医药, 2016, 23(32): 4-6转10.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2016/V23/I32/4

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