关键词: Alb, 慢病毒载体, HBEGF, copGFP

Abstract: 【Abstract】〓Objective〓To generate lentiviral vector carrying HBEGF gene under the control of liver-specific albumin (Alb) promoter. Methods〓Firstly, the fragment of Alb-enhancer/promoter was amplified from pAlb-Cre-GH/BS, and subsequently subcloned into the Not I site in the polylinker of pTRECK6-GFP by In-Fusion cloning to generate pAlb-TRECK6..Secondly,.the fragment of Alb-TRECK6 was amplified from pAlb-TRECK6,.and then inserted into pCD823A-1 at the Xba I/BamH I sites by In-Fusion cloning, designated pLV-ATTG, as verified by restriction enzyme digestion and sequencing. According to the standard protocol from Invitrogen,.lentiviruses were produced and then infected BEL-7402 cells,.followed by copGFP assay under inverted fluorescence microscope, and detecting the expression of HBEGF transgene by qRT-PCR. Results〓the data from enzyme digestion and DNA sequencing demonstrated that pLV-ATTG was successfully constructed..Cells infected with LV-ATTG displayed the green fluorescence under inverted fluorescence microscope, and the increased expression of HBEGF detected by qRT-PCR. Conclusion〓The lentiviral vector harboring HBEGF gene under the control of liver-specific Alb promoter was successfully constructed,.which will lay a solid foundation for further research.

Key words: HBEGF, CopGFP, Lentivirus vector, Alb