Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene
- VernacularTitle:卡氏肺孢子表面糖蛋白siRNA表达载体的构建和鉴定
- Author:
Lini WANG
- Publication Type:Journal Article
- Keywords:
Pneumocystis carinii;
Major surface glycoprotein;
siRNA;
Expression plasmid
- From:
Journal of Chongqing Medical University
2003;0(05):-
- 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.Methods:Short hairpin RNA(shRNA) oligonucleotide targeting PC MSG gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6+1 after annealing.the recombinant plasmid, pPC-MSG,transgormed into E.coil.TOP10 and amplified,was digested by restriction endonucleases HindⅢand EcoRⅠand identified by gelelectrophoresis 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-MSG was constructed successfully.
