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Abstract

BACKGROUND:

Studies have confirmed that vascular endothelial growth factors (VEGF) play an important role in the course of normal liver regeneration, while, there are very few reports on the same role in the cirrhotic liver nationally and internationally.

OBJECTIVE:

To construct the lentivirus vector containing human VEGF165 gene, and to examine the expression of VEGF165 in normal rat liver cells after transfecting BLR 3A rat liver cells, in order to lay a foundation for the study about the effects of VEGF165 on the regeneration power of remnant liver tissue after hepatectomy in the rats with cirrhosis.

METHODS:

The VEGF165 gene was cloned to a lentiviral expression vector by recombinant DNA technology. The positive clones were screened, and lentiviral packaged systems (ViraPowerTM Packaging Mix) were co-transfected to package virus in 293T cells by lipofection with target gene plasmid. Real-time PCR technique was used o test the titer of pLenti6/V5-D-TOPO-VEGF165. The BLR 3A rat liver cells were transfected by pLenti6/V5-D-TOPO-VEGF165 and the expression of VEGF165 mRNA and protein was detected by reverse transcriptase-PCR and Western blot technique.

RESULTS AND CONCLUSION:

The lentivirus vector containing VEGF165 was successfully constructed. Virus titer could reach as high as 1.18×107 VP/mL. The transfer efficiency of green fluorescent protein was more than 80% in the BLR 3A rat liver cells after transfection with pLenti6/V5-D-TOPO-VEGF165 for 72 hours. Reverse transcriptase-PCR and Western blot results showed that the VEGF165 gene expression of the transfected group was positive. Lentivirus vector pLenti6/V5-D-TOPO-VEGF165 could transfect BLR 3A rat liver cells effectively, and VEGF165 gene was expressed after transfection.

RESULTS AND CONCLUSION:

Yi C, Wang XY, Li HJ, Wang H. Construction and expression of human vascular endothelial growth factor 165 gene lentiviral expression vector. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(11): 1927-1932. [http://www.crter.cnhttp://en.zglckf.com]

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