Amplification and cloning of functional fragment of hTERA cDNA

Objective: To assay the telomerase activity level in human laryngocarcinoma cell line hep 2,and to amplify the functional fragment of hTERT (human telomerase reverse transcriptase) cDNA in hep 2.Methods:TRAP ELISA (telomere repeat amplification protocol enzyme linked immunosorbent assay) was used to assay the level of telomerase activity,and RT PCR,to amplify the target fragment.Then,restriction analysis of EcoR Ⅰ and BamH Ⅰ,PCR identification,and DNA sequence analysis were carried out to verify the recombinant plasmids.Results:The telomerase activity level was high in hep 2;the clone carrying the target fragment of hTERT cDNA was obtained.Conclusion:hTERT cDNA can be amplified by RT PCR from hep 2 with high telomerase activity level. [