Construction and expression of mouse liver-specific expression vector Kbpala/Alb-STBXATP7B / 中国病理生理杂志

ABSTRACT

AIM: To construct the liver-specific transgene vectors encoding wild-type as well as most common disease mutant STBXATP7B cDNA under the control of mouse albumin promoter and to explore their expression. METHODS: Two Kbpala/Alb-STBXATP7B mutants containing the Arg778Leu and His1069Gln mutations were constructed using site-directed mutagenesis system plus site-subcloning technique. The vectors expressed wild-type and mutants of human STBXATP7B in mouse liver cells were obtained and transiently transfected into BRL and BHK cell lines. Western blotting analysis was utilized to detect the expression of human STBXATP7B. RESULTS: Enzyme analysis and sequencing analysis confirmed that the target genes were STBXATP7B and in right position. The results of Western blotting showed that the gene products were expressed specifically in liver cells. CONCLUSION: The Kbpala/Alb-STBXATP7B vectors were constructed successfully and the liver-specific expression of human STBXATP7B proteins were conformed.