Construction And Expression Of HTERT In Eucaryotic Expression Vector

Background and Objectives:Telomerase is a ribonucleoprotein complex with terminal transcription activity. it is expressed in the vast majority of human malignant tumors. Telomerase is composed of an RNA component (hTR), the telomerase-associated protein TEPl, and a catalytic protein subunit (hTERT). 80-90%tumor telomerase activity is positive and hTERT-RNA (mRNA) is corresponded with activation of telomerase, it is shown that the telomerase catalytic subunit the key determinant plays an important role in human malignant tumor tissue telomerase activity. Normal tissue \cell telomerase activity is negative except for stem cell\stimulated lymphocyte\gonad cell, reconstruction of telomerase activity is the important molecular event in oncogenesis, is also the important target in anti-tumor therapy. Recently, investigation of activating and repressive mechanisms of the hTERT transcription in cancer has become an area of intense interest in cancer research. In order to explore the feasibility of hTERT gene as a tumor vaccine, we insert genomic fragment of hTERT into the pGEM-T Easy vector, subcloned into eukaryotic expression vector pEGFP-C3. After identified, the recombinant was transfected into Mouse fibroblast cell line NIH3T3 cell by Lipofectamine method, observe the effect of transfectant. It makes the foundation for anti-tumor immunity therapy associated with hTERT.