Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface glycoprotein gene's upstream conserved sequence
- VernacularTitle:卡氏肺孢子主要表面糖蛋白UCS基因siRNA表达载体的构建和鉴定
- Author:
Yan TAN
- Publication Type:Journal Article
- Keywords:
Pneumocystis carinii MSG-UCS;
siRNA;
Expression vector
- From:
Journal of Chongqing Medical University
1986;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct and identify a plasmid vector of short interfering RNA(siRNA)on pneumocystis carinii(PC) major surface glycoprotein(MSG)gene's upstream conserved sequence(UCS).Methods:Short hairpin RNA(shRNA) oligonucleotide targeting PC MSG UCS gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6+1 after annealing.The recombinant plasmid,PC-UCS,transformed into E.coil.TOP10 and amplified,was digested by restriction endonucleases HindⅢand EcoRI and identified by gel electrophoresis and DNA sequencing.Results:Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion:The siRNA plasmid,pPC-UCS,was constructed successfully.
