Construction and identification of pSilencer~(TM) 3.1-H1 neo mdr1 short hairpin RNA expression vectors

Objective:To study the construction of pSilencerTM 3.1-H1 neo mdr1 short hairpin RNA expression vectors.Methods:Two different shRNA targets desigened to be homologous to the P-glycoprotein(P-gp)encoding mdr1 mRNA consensus sequence,were annealed and ligated into the BamHⅠand Hind Ⅲ site of linearized pSilencerTM 3.1-H1 neo vector.The recombinant named as pSilencerTM3.1-H1 neo mdr1-A and mdr1-B shRNA expression plasmids was identificated by restrictive enzyme digestion and sequencing.Results:The fragments of 66 bp and 4.3 kbp were shown after digestion by BamHⅠand Hind Ⅲ and agarose gel electrophoresis.The DNA sequences of pSilencerTM3.1-H1 neo mdr1-A and mdr1-B shRNA expression plasmids were proved to be identical to the data of mdr1 in Genebank.Conclusions:pSilencerTM3.1-H1 neo mdr1 shRNA expression vectors has been constructed successfully.