Cloning of Tumor Metastasis-suppressor Gene KiSS-1 and Construction of its Eukaryotic Expression Vector

Objective To clone the tumor metastasis suppressor gene Kiss-1 from normal human urinary bladder tissue which doesn't contain the signal peptide,and to construct its eukaryotic expression vector.Methods Total RNA was extracted from human urinary bladder tissue and the opening reading frame of KiSS-1 cDNA was isolated through RT-PCR as PCR template.Then,cloned it into the eukaryotic expression vector pcDNA3.1(+).The expression vector of pcDNA3.1(+)/KiSS-1 was constructed successfully.Results The recombinant plasmid was identified by sequencing analysis,PCR and restriction enzyme digestion.It was testified that the target gene fragment was successfully cloned into the recombinant plasmid pcDNA3.1(+)as expected.Conclusions The successful construction of the recombinant plasmid pcDNA3.1(+)/KiSS-1 will be benefit to the further study in the protein expression and the protein's function.