Cloning of human bone morphogenetic protein-2 gene and construction of recombinant adenoviral vector
- VernacularTitle:人骨形成蛋白-2基因的克隆及其重组腺病毒载体的构建
- Author:
Li ZHANG
;
Ning MA
;
Wenxin GAO
;
Ming ZHANG
- Publication Type:Journal Article
- Keywords:
bone morphogenetic proteins;
genetic vectors;
genes;
transfection
- From:
Journal of Jilin University(Medicine Edition)
2006;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone the human bone morphogenetic protein-2(hBMP2) gene cDNA and construct its replication-deficient type 5 adenoviral vector.Methods Using osteoblastic sarcoma tissue to extract cell total RNA,hBMP-2 cDNA was amplified by RT-PCR.The hBMP-2 cDNA was then inserted into cloning vector pGEM-T.The recombinant plasmid was screened and the recombinant plasmid which was confirmed by restriction enzyme,PCR and sequence analysis was gotten.To construct the recombinant adenoviral vector,the BMP-2 cDNA was subcloned into a shuttle vector and the recombinant pShuttle-BMP2 vector was obtained.It was subcloned into Adeno-X adenoviral genomic DNA and the pAd-BMP2 DNA was obtained.Results A length of 1 200 bp BMP-2 cDNA was cloned,and it was subcloned into adenoviral vector,the recombinant adenovirus was constructed successfully.After measuring the titre of virus(1?1011 nfu?L-1),the target gene was evaluated by PCR.Conclusion The hBMP-2 adenoviral is obtained.The successful construction of pAd-BMP2 may provide us useful target for the further research.
