Construction of Polypetide N-acetylgalactosaminyltransferase 2 Eukaryotic Expression Vector and Expression in SHG44 Cells

Objective: To construct the eukaryotic expression plasmid pcDNA3/ppGalNAc-T2 and transfect it into SHG44 cells for exploring the effect and function of genesis,development and metastasis in cancer. Methods:With PCR the target DNA fragment enconding ppGalNAcT2 was amplified from cloning plasmid pDONR201-T2 and subcloned to the eukaryotic expression plasmid pcDNA3.1,then transfected the recombinant plasmid into SHG44 cells,and detected the transcription of ppGalNAcT2 in transfected cells by RT-PCR.Results: Certificated by restriction enzymes digestion and DNA sequencing,the eukaryotic expression plasmid pcDNA3.1-T2 was constructed successfully and the T2 mRNA was detected by RT-PCR in transfected cells.Conclusion:The eukaryotic expression plasmid pcDNA3.1-T2 was successfully constructed and the recombinant plasmid was successfully transfected into SHG44 cells,which will lay the foundation for the study of its function.