Construction of rat bowel transforming growth factor beta1 recombinant adenovirus vector.
- Author:
Ji-peng LI
1
;
Yi HUANG
;
Wei-zhong WANG
;
Guang-long DONG
;
Jian-jun DU
;
Dong-li CHEN
;
Gang JI
;
Ji LIU
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Animals; Cells, Cultured; Cloning, Molecular; Dendritic Cells; Gene Expression; Genetic Vectors; Intestine, Small; metabolism; Rats; Recombination, Genetic; Transfection; Transforming Growth Factor beta1; genetics
- From: Chinese Journal of Gastrointestinal Surgery 2008;11(1):76-79
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a recombinant adenovirus vector which regulates the expression of rat transforming growth factor beta (TGF-beta1).
METHODSTotal RNA was extracted from F344/N rat small intestine pre-treated with Con A to clone TGF-beta1. pTRE-shuttle vector was used as mediator to ligate TGF-beta1 gene and backbone of replication-incompetent adenoviral vector. The constructed recombinant adenovirus contained tetracycline-responsive element which could regulate the expression of inserted genes. After identification, the desired recombinant adenovirus was packaged in HEK 293 cells. Supernatant of high titer adenovirus was collected to detect the TGF-beta1 gene expression by green fluorescent protein(GFP).
RESULTSThe constructed recombinant adenovirus was identified by restriction endonucleases cutting, sequencing, PCR and GFP examination.
CONCLUSIONRat TGF-beta1 recombinant adenovirus is established successfully, which provides material and evidence for further research of dendritic cell (DC) modified by TGF-beta1 to induce immune tolerance in rat heterotopic small bowel transplantation.
