Construction and identification of vectors expressing short hairpin RNA targeting mouse STAT4 and STAT6 gene
- VernacularTitle:小鼠STAT4,STAT6基因shRNA真核表达质粒的构建及鉴定
- Author:
Qin YANG
- Publication Type:Journal Article
- Keywords:
Asthma;
Signal transducers and activators of Transcription;
RNAi;
shRNA;
Plasmid
- From:
Journal of Chongqing Medical University
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct short-hairpin RNA(shRNA)expressing vectors aimed at mouse signal transducers and activators of transcription-4/6(STAT4/STAT6). Methods: Target sequences in the middle of STAT4 and STAT6 gene were selected, and then the complementary 65 mer oligonucleotides of each sequence were synthesized. After annealed,the inserts were ligated into the linearized pGenesil-3 plasmid. The plasmids were transformed into DH5? bacteria to amplify and then purified. GAPDH plasmid was constructed as a positive control and targeting none gene was served as a negative control. These purified plasmids were identified by gel electrophoresis after being cut with restriction enzyme and sequencing. Results: The results of gel electrophoresis and sequencing showed that the plasmids were identical with the positive control, and the sequences were identical with what we have inserted in. Conclusion: The vectors which can express siRNA hairpin aimed at STAT4/STAT6 gene have been constructed successfully and it lays the groundwork for the further study.
