Characterization of hepatocyte growth factor expressed by baculovirus.
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.
Lee
H S
HS
Department of Internal Medicine, National Taiwan University, Taipei, R.O.C.
Huang
G T
GT
Sheu
J C
JC
Chiou
L L
LL
Lai
M Y
MY
Chen
D S
DS
Lee
S C
SC
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D004285:Dogs; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D007668:Kidney; D008099:Liver; D008113:Liver Neoplasms; D008168:Lung; D051379:Mice; D008970:Molecular Weight; D009036:Moths; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D017228:Hepatocyte Growth Factor