Construction of a recombinant adenovirus vector for interferon-gamma-inducible protein 10 and the adenovirus preparation.
- Author:
Zi-yun SHAO
1
;
Zhi-feng LIU
;
Yi PENG
;
Jia XU
;
Peng DENG
;
Yong JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Cell Line; Chemokine CXCL10; genetics; metabolism; Cloning, Molecular; Defective Viruses; genetics; Genetic Vectors; genetics; Green Fluorescent Proteins; genetics; metabolism; Humans; Recombinant Fusion Proteins; genetics; metabolism; Transfection; Virus Cultivation; methods
- From: Journal of Southern Medical University 2006;26(11):1552-1555
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a recombinant adenovirus vector for expressing interferon-gamma-inducible protein 10 (IP-10) by homogenous bacterial recombination.
METHODSIP-10 gene was cloned into the shuttle plasmid pAdTrack-CMV that contained the coding sequence of enhanced green fluorescent protein (EGFP). The shuttle plasmid was then transformed into E. coli BJ5183 with pAdEasy-1 vector by chemical transformation. The recombinant adenovirus vector pAd/IP-10 was identified by enzyme digestion with Pac I and the linearized plasmid was transfected into HEK293 cells.
RESULTSThe positive clones were identified with enzyme digestion and polymerase chain reaction (PCR) and were further verified by DNA sequencing. The recombinant adenovirus of high titration was obtained after transfection and packaging in HEK293 cells.
CONCLUSIONA recombinant adenovirus vector for expression of IP-10 has been constructed successfully and high-titer active adenovirus is obtained for functional study of IP-10 protein.
