Lee H S, Huang G T, Sheu J C, Chiou L L, Lai M Y, Chen D S, Lee S C
Department of Internal Medicine, National Taiwan University, Taipei, R.O.C.
Biochem Biophys Res Commun. 1993 Dec 15;197(2):591-8. doi: 10.1006/bbrc.1993.2520.
To express recombinant hepatocyte growth factor (HGF), we cloned the cDNA by polymerase chain reaction. Functional recombinant HGF was expressed in insect cells by a baculovirus expression system. The recombinant HGF is a mixture of a single-chain precursor and a two-chain heterodimer. Under non-reducing conditions, they migrate together as a single band with MW 67 kDa on Western blots. Under reducing conditions, the precursor migrates at 91 kDa, alpha-chain at 64 kDa and beta-chain at 34 kDa. About 25% of the expressed HGF is in the expression medium, which does have scatter activity on HepG2 and MDCK cells and proliferation activity on BN CL.2 cells (a mouse hepatocytes cell line) in a dose-dependent manner.
为了表达重组肝细胞生长因子（HGF），我们通过聚合酶链反应克隆了cDNA。功能性重组HGF通过杆状病毒表达系统在昆虫细胞中表达。重组HGF是单链前体和双链异二聚体的混合物。在非还原条件下，它们在蛋白质免疫印迹上作为一条分子量为67 kDa的单带一起迁移。在还原条件下，前体迁移至91 kDa，α链迁移至64 kDa，β链迁移至34 kDa。约25%的表达的HGF存在于表达培养基中，其对HepG2和MDCK细胞具有散射活性，对BN CL.2细胞（一种小鼠肝细胞系）具有增殖活性，且呈剂量依赖性。
