Construction and significance of the short hairpin RNA recombinant plasmid targeting M2 gene of respiratory syncystial virus
- VernacularTitle:靶向呼吸道合胞病毒M2基因pshRNA载体质粒的构建及意义
- Author:
Yuxia CUI
- Publication Type:Journal Article
- Keywords:
Respiratory syncystial virus;
M2 gene;
Short hairpin RNA;
Recombinant plasmid
- From:
Journal of Chongqing Medical University
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct the recombinant plasmid targeting M2 gene of respiratory syncystial virus(RSV) for further study of RNA interference(RNAi) technology in inhibiting RSV infection.Methods: According to the designing principles of shRNA,The M2 gene of RSV was selected as the target gene.A 19bp reverse repeated sequences with 9bp spacer were designed and synthesized.The complement strain were obtained by annealing and inserted into the vector pgenesil-1 containing green fluorescenet protein(GFP) sequence and U6 promotor.Finally the recombinant plasmid were identified by enzyme digesion and DNA sequencing,then transfected into HEP2 cells and observed by fluorescence microscope.Results: The recombinante plasmid targeting the mRNA of RSV M2 gene was successfully constructed and the good efficents of transfection were found by fluorescence microscope.Conclusion: The successful construction of the pshRNA recombinant plasmid targeting the M2 gene of RSV will help futher study on application of the RNAi technology to anti-RSV infection.
