Construction of Recombinant Adenoviral Vector Carrying the Human BMP-2 Gene with the Technology of in vitro Ligation
- VernacularTitle:人骨形成蛋白-2基因腺病毒表达载体构建方法的优化
- Author:
Lei WANG
;
Xun QU
;
Fengcai WEI
;
Jinbo FENG
;
Shanzhen SUN
;
Shaohua LIU
;
Meixiang YANG
- Publication Type:Journal Article
- Keywords:
adenovirus;
BMP-2;
gene recombination;
in vitro ligation
- From:
Chinese Journal of Cancer Biotherapy
1995;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To provide an efficeut protocol for constructing recombinant adenovirus, an in vitro ligation was used instead of homologous recombination. Methods: Gene BMP-2 was ligated into pShuttle 2 vector ( pShuttle 2-BMP-2 ) and then fragment containing BMP-2 gene and promoter pcmvie excised by PI-SCe Ⅰ and Ⅰ-Ceu Ⅰ endonuclease. The fragment was further combined with adenovirus vector (Adeno-X-BMP-2) , which was finally linearized with Pac Ⅰ and trans-fered to HEK293 to package adenovirus particles. Results: Both PCR assay and restiction analysis showed that the recombined rectors pShuttle2-BMP-2 and Adeno-X-BMP-2 contains the target BMP-2 gene. THe packaged adenovirus was also i-dentified by PCR assay with specific primers for BMP-2. Conclusions: The BMP-2 incorporated recombinant adenovirus was obtained and this laid a foundation for further study on BMP-2 mediated gene therapy. The in vitro ligation method de-scinbed here for constructing recombined adenovirus was more efficient than traditional homologous recombination.
