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

Objective: To study the construction of pSilencer^TM 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 BamHⅠ and Hind Ⅲ site of linearized pSilencer^TM 3.1-H1 neo vector.The recombinant named as pSilencer^TM3.1-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 BamHⅠ and Hind Ⅲ and agarose gel electrophoresis.The DNA sequences of pSilencer^TM3.1-H1 neo mdr1-A and mdr1-B shRNA expression plasmids were proved to be identical to the data of mdr1 in Genebank.Conclusions: pSilencer^TM3.1-H1 neo mdr1 shRNA expression vectors has been constructed successfully.