Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface gene / 重庆医科大学学报

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.