Abstract:

Abstract:Objective To construct eukaryotic expressing vector of PTEN gene and to express the gene in the Hep-2 cell line and to clarify theraputic effect  of PTEN gene in the Hep-2 cell line,and  may laid foundation for further  study of PTEN gene treatment of laryngeal carcinoma.Methods PTEN gene was amplified from human normal laryngeal mucosa by RT-PCR and the fragment of the cDNA was cloned into the PTEN-Teasy vector.After the selection of the positive clone by DNA sequencing and restriction enzyme analysis,the fragment of the cDNA was cloned into the eukaryotic expressing vector of pEGFG-C1.The recombinant plasmid transfected into Hep-2 cell line and the gene transient expression was observed.with Western blotting.Results  A  1 400 bp DNA fragment was amplified with RT-PCR.The sequence analysis showed it was consistant with the sequence of PTEN gene in GenBank.Western blotting analysis provided strong evidence that PTEN gene was expressed successfully in transfected Hep-2 cell line.Conclusion The eukaryotic expressing vector of PTEN gene is constructed and PTEN protein is induced to express successfully.

Key words: anti-oncogene;PTEN gene;gene cloning; , gene expression