Expression of angiostatin using DNA-based semliki forest virus replicon.
Angiogenesis is recognized as a critical factor in the growth of tumor cells and plays a key role in the tumor metastasis. Recent studies for antiangiogenic substances are getting popular. The angiostatin, one of the antiangiogenic substances, leads to the increased apoptosis of the tumor cells by inhibiting the neovascularization of the tumor. The angiostatin was identified as the internal fragments of the plasminogen which has no antiangiogenic activity. By hydrolysis of the plasminogen, the angiostatin can be produced. In this study, we constructed the SFV-derived DNA vector by employing the cytomegalovirus immediate early enhancer/ promoter (CMV). This vector makes it possible to transfect the cells with DNA without the in vitro transcription process. The C-myc epitope and polyhistidine residue sequences were placed in downstream of the angiostatin gene to make it eligible to detect the expressed protein. The murine Ig k-chain V-J2-C signal sequence was placed in upstream to secrete the expressed protein from the cells. We confirmed the expression of angiostatin in the BHK-21 cells using DNA-based SFV replicon.
Choi
Yong-Soo
YS
College of Veterinary Medicine, Chungnam National University, Daejon, 305-764, Korea.
Lee
Jong-Soo
JS
Choi
Young-Ki
YK
Shin
Kwang-Soon
KS
Kim
Hyun-Soo
HS
Kim
Chul-Joong
CJ
D043168:Angiostatins / Q000032:analysis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers; D015967:Gene Expression Regulation, Viral; D007150:Immunohistochemistry; D007668:Kidney; D010957:Plasmids; D012093:Replicon / Q000235:genetics*; D012672:Semliki forest virus / Q000235:genetics*; D014162:Transfection
D017931:DNA Primers; D043168:Angiostatins