Eukaryotic Expression of 14-3-3 Signal Protein from Schistosoma Japonicum (Chinese Mainland Strain)

Objective Sj14-3-3 signal protein gene from cDNA of Schistosoma Japonicum adult worms was amplified and subcloned into eukaryotic expression vector pcDNA3.1(+) for a new vaccine candidate of S. Japonicum. Methods The total RNA of S. Japonicum was extracted to prepare for cDNA by RT-PCR, from which the Sj14-3-3 encoding gene was amplified. The DNA fragment was subcloned into eukaryotic expression vectors pcDNA3.1(+) following the insertion and amplification in pGEM-T. The recombinant plasmid was transfected into COS-7 cells and identified by Western blotting. Results The size of RT-PCR product was approximately 760 bp and the inserts of pcDNA3.1(+)-Sj14-3-3 and pGEM-T-Sj14-3-3 digested with BamH I and Xho I were all the same in length as RT-PCR product. The expression was confirmed by monoclonal antibody against Sj14-3-3.Conclusion The recombinant DNA with Sj14-3-3 could be expressed in COS-7 cells with a molecular weight of 29 kD.