Construction of eukaryotic expression plasmid of hTERT.

Objective To construct human telomerase reverse transcriptase (hTERT)expression plasmid,which is helpful to establish immortalized cell lines and begin clinical cell transplantation in experimental study and treatment of disease. Methods The hTERT full-length cDNA sequence was amplified by RT-PCR using a human fetal brain cDNA library. The amplified product was ligated with the pGEM-T vector,which was followed by DNA sequencing. Results Eukaryotic expression plasmid of pGEM-T-hTERT was identified by the digestion of restriction endonuclease. The DNA sequence of hTERT fragment was compared with that in GenBank for homology. The homology was 98.87 % in nucleotide acid,and 99.67 % in amino acid. Conclusion The eukaryotic expression plasmid of hTERT has been successfully constructed.