Amplification and clone of the gene coding for proteinase cathepsin L2 in Schistosoma japonicum in mainland China

Aim In order to provide the basis for further study on the function of proteinase cathepsin L2 in Schistosoma japonicum amplifying the gene coding for SjCL2 in vitro and cloning the fragment into the eukaryotic expression vector pcDNA3.Methods Total RNA was isolated from adult worms of S.japonicum in China mainland using TRIZOL.The coding region gene of SjCL2 was amplified by RT-PCR.The fragment from RT-PCR was cloned into eukaryotic expression vector pcDNA3 via BamHⅠ and XhoⅠ sites.The resulting construct was determined by restriction and PCR analysis methods.Results The coding region of SjCL2 gene,which was about 1kb,was specifically amplified by RT-PCR.The expression plasmid pcDNA-SjCL2 contained the amplified fragment,which was validated by restriction and PCR analysis.Conclusion The results show that the length of amplified fragment is identical with the predicted one,and the SjCL2 eukaryotic expression plasmid has been successfully constructed.