Construction of Retroviral Recombinant Vectors Carrying Sense- and Antisense-hGR? cDNA by an Improved Ligasing Protocol
- VernacularTitle:改进方案构建人糖皮质激素受体?基因正、反义重组逆病毒载体
- Author:
Min ZHU
;
Qingnan HE
;
Gang ZHOU
- Publication Type:Journal Article
- Keywords:
Human glucocorticoid receptor ?;
Ligase;
Antisense RNA
- From:
Journal of Chinese Physician
2001;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant expression vector carrying human glucocorticoid receptor ?(hGR?) cDNA to provide experiences for cloning of large DNA fragments and offer experimental materials for studying the function and medchanism of hGR? by antisense nucleic acid technique. Methods hGR? cDNA was amplified from an expressional phage library of human white cells by L-PCR, and cloned into pGEM-Teasy vector. Subsequently, the hGR? cDNA fragment digested from the recombinant T vector was sub-cloned into retroviral vector pLXSN in sense and antisense orientations, respectively through an improved ligasing protocol. Results Restriction endonuclease analysis and sequencing showed that cloning and sub-cloning of hGR? were successful. Conclusion Retroviral recombinant vectors carrying sense-and antisense-hGR? cDNA were successfully constructed.
