J Vet Sci.  2002 Mar;3(1):41-45.

Expression of Angiostatin Using DNA-Based Semliki Forest Virus Replicon

• Choi YS
• Lee JS
• Choi YK
• Shin KS
• Kim HS
• Kim CJ

Abstract

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 kappa-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.

MeSH Terms

Angiostatins/analysis/*genetics
Animals
Base Sequence
Cell Line
Cricetinae
DNA Primers
Gene Expression Regulation, Viral
Immunohistochemistry
Kidney
Plasmids
Replicon/*genetics
Semliki forest virus/*genetics
Transfection