Construction of replication deficient recombinant adenovirus of monocyte chemoattractant protein-1
- VernacularTitle:表达人单核细胞趋化蛋白-1基因重组腺病毒的制备
- Author:
Heping PENG
;
Yongguo LI
;
Hao GUAN
;
Puning WAGN
;
Qicai LIU
- Publication Type:Journal Article
- Keywords:
Monocyte chemoattractant proteins/biosynthesis;
Adenoviridae;
DNA,recombinant
- From:
Journal of Chinese Physician
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct replication deficient recombinant adenovirus of human monocyte chemoattractant protein-1(MCP-1) by homologous recombination.Methods The cDNA of MCP-1 gene was obtained from human liver tissue by using RT-PCR,and was subcloned into a transfer plasmid pAdTrack-CMV.The linearized recombinant transfer plasmid pAdTrack-CMV-MCP-1 was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral plasmid.The recombinant adenoviral plasmid was linearized and then transfected into HEK293 packing cells to produce virus particles.The recombinant adenovirus was detected by using PCR.Results The recombinant adenoviral plasmid was successfully established and confirmed by restriction endonuclease digestion.The expression of green fluorescent protein(GFP) was observed on the 5th day after transfection.The fragment of MCP1 gene was amplified by PCR.Conclusion The achievement of recombinant adenoviral plasmid and recombinant adenovirus of MCP-1 lay a foundation for further investigation of the function and application of MCP-1.
