Cloning of human KiSS-1 gene and construction of its eukaryotic expression vector

Objective: To clone the metastasis suppressor gene KiSS-1 from human normal human pancreas tissue and construct its eukaryotic expression vector. Methods: Total RNA was extracted from human pancreas tissue. The opening reading frame of KiSS-1 cDNA was isolated by using RT-PCR, and cloned into eukaryotic expression vector pcDNA3. The expression plasmid pcDNA3/KiSS-1 was constructed. Results: The recombinant plasmid was identified with PCR, digestion of internal restriction enzyme, and sequencing. The nucleotide sequence isolated from the recombinant plasmid pcDNA3/KiSS-1 was the same as expected. Conclusion: The successful construction of the recombinant plasmid pcDNA3/KiSS-1 will be benefit to further study in the protein expression and the antibody production.