Construction of retroviral vector inserted SV40 large T antigen gene and its expression in rat hepatocytes
- VernacularTitle:SV40LT抗原基因逆转录病毒载体的构建及其在肝细胞中的表达
- Author:
Yi ZHANG
;
Weijia SUN
;
Xuejun LAO
;
Yangde ZHANG
- Publication Type:Journal Article
- Keywords:
LIVER NEOPLASMS/genet;
SV40 LARGE T ANTIGEN GENE
- From:
Chinese Journal of General Surgery
2001;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the retroviral vector inserted SV40 large T antigen gene and transfect it into rat hepatocytes, analyze the status of SV40 large T antigen gene expression in rat hepatocytes, and to establish important basis for clinical hepatocytes transplanation. Methods Retroviral vector inserted SV40 large T antigen gene was(constructed) by DNA recombinant techniques in vitro, then the combinant vector was determined with enzyme(digestion) and sequencing and was transfected into the PA317 cell lines by liposome mediation and screened(anti-G4)18 positive clones. The viral titer was determined with the NIH3T3. After transfected into separated and purified rat primary hepatocytes, the SV40 large T antigen gene expression was detected by PCR and immunohistochemical (methods). Results (1)SV40 large T antigen gene fragment was inserted into retroviral vector in sense orientation. (2)The titer of pseudovirion packed by PA317 cell lines was 1.3?10~6CFU/ml. (3)SV40LT antigen gene was(integrated) into rat primary hepatocytes and its expression in transfected cells at 24 hour was higher than 96 hour (P
