Cloning and expression of human DcR3 molecule in COS-7 cells
- VernacularTitle:人DcR3cDNA克隆及其在COS-7细胞中的表达
- Author:
Huaizhi WANG
;
Wanling LI
;
Jiahong DONG
;
Bing NI
;
Man JIANG
;
Yuzhan WU
- Publication Type:Journal Article
- Keywords:
DcR3;
COS-7 cells;
gene expression
- From:Journal of Third Military Medical University
2003;0(17):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone ORF of DcR3 gene and insert it into eukaryotic expression vector and express it in COS-7 cells. Methods Encoding sequence of human DcR3 gene was cloned by PCR and sequenced. The sequenced ORF was cloned into eukaryotic expression vector pAAV-IRES-hrGFP to construct recombinant plasmid. COS-7 cells were transfected with recombinant plasmid by lipofectamine2000. Expression of recombinant DcR3 gene was verified by Western blotting and confocal microscopy. Results A 1 000-bp gene segment was obtained by PCR and inserted into pAAV-IRES-hrGFP to construct recombinant plasmid. The gene segment was proved to be encoding sequence of human DcR3 gene by sequencing. DcR3 expression in COS-7 cells was verified by Western blotting and confocal microscopy. Conclusion DcR3 gene was successfully cloned and expressed in COS-7 cells.
