Construction and application of luciferase reporter plasmid drived by ALB promotor
AIM: To construct a luciferase reporter drived by ALB promoter and apply it to investigate the effect of VEGF in the differentiation of hepatic progenitor cell. METHODS: ALB promoto was amplified from genomic DNA and then cloned into pBGLuc multipl cloning sites to construct the luciferase reporter plasmid pBGLuc-ALB. The plasmid was verified by agarose gel electrophoresis, DNA sequencing, and function test. The stable hepatic progenitor cell line E14.5-ALB-LUC, expressing luciferase drived by ALB promote was treated with adenovirus VEGF. The differentiation of the cell line E14.5-ALB-LUC was evaluated by the expression of luciferase and confirmed by RT-PCR and PAS staining. RESULTS: Agarose gel electrophoresis, DNA sequence and t function test verified the successful construction of the plasmid pBGLuc-ALB. Compared with control group, the expression of luciferase in the hepatic progenitor cell line E14.5-ALB-LUC treated with adenovirus VEGF increased. At the same time, the expression of early marker DLK, CK19 more clearly decreased while ALB are opposite in experimental group. The morphological change mainly are increased glycogenesis. CONCLUSION: The recombinante plasmid pBGLuc-ALB drived by ALB promoter has been successfully constructed and could be used to evaluate the differentiation of hepatic progenitor cell. VEGF can induce the differentiation of hepatic progenitor cell.
NETL
NSTL
万方数据
维普
