血管内皮生长因子原核表达载体的构建及表达

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Abstract Objective To construct a prokaryotic expression vector containing vascular endothelial growth factor (VEGF) and induce to express protein, it may lay a foundation for the follow-up study of VEGF. Methods Amplified The target fragment was amplified by reverse transcription polymerase chain reaction (RT-PCR) and the DNA fragment was cloned into pET-30a(+) vector with histidine tag. The recombinant plasmid was identified by polymerase chain reaction (PCR) and sequencing. The expression of recombinant plasmid was induced by isopropylthio-β-D-galactoside (IPTG) and identified by Western blot. Results The right size of the fragment was amplified from HepG2 cells. The recombinant plasmid pET-30a-VEGF was confirmed by PCR and DNA sequencing. Coomassie brilliant blue staining and Western blot results showed that the molecular weight of the fusion protein induced by IPTG was correct and the band was single. Conclusion The prokaryotic expression plasmid of pET-30a-VEGF is successfully constructed, in addition, it might lay the foundation for development of HCC diagnostic and treatment research.

Key words： Vascular endothelial growth factor Hepatocellular carcinoma Gene cloning Prokaryotic expression

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	PANG Lijun1 LIU Daojie2 LIN Minghua1 LIU Kai1 LIU Xiaoni1 CHEN Dexi1

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PANG Lijun1 LIU Daojie2 LIN Minghua1 LIU Kai1 LIU Xiaoni1 CHEN Dexi1. Construction and expression of prokaryotic expression vector of vascular endothelial growth factor[J]. 中国医药导报, 2020, 17(23): 4-7.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2020/V17/I23/4

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