Construction and Identification of CD147 siRNA Expression Vector / 中华皮肤科杂志

ABSTRACT

Objective To construct CD147 siRNA expression vector in o rder to analyze the role of CD147 in the invasion and metastasis of tumors deriv ed from skin.Methods According to CD147 cDNA sequence in the Genebank,a pair of 64-nt oligonucleotides,each containing the sites of restriction endonucleas e at both ends,were designed and synthesized.Oligonucleotides were annealed an d ligated with linearized pSUPER by T4DNA ligase.The recombinants (named pSUPER /CD147 siRNA ) were finally sequenced and identified by PCR and restriction end onuclease digestion.Results CD147 siRNA expression vector was successfully co nstructed and identified by double endonuclease digestion.Sequence analysis of inserted fragment revealed the same sequence as synthesized siRNA oligonucleotid es.Conclusions CD147 siRNA expression vector has been successfully constructed,which paves the way for studying the role of CD147 in the invasion and metasta sis of tumor cells derived from skin as well as in tumor therapy.