Abstract:

BACKGROUND: Human telomerase reverse transcriptase is an important component of telomerase. The display on telomerase activity results in anti-telomere shortening or cell senescence.
OBJECTIVE: To construct an eukaryotic expressing vector containing the C-terminal amino acid 936-1132 of human telomerase reverse transcriptase; to express the constructed vector in human embryonic kidney 293T cells; to lay the foundation for the research of intracellular localization.
METHODS: The C-terminal fragment of human telomerase reverse transcriptase containing amino acids 936-1132 and enzyme sites was amplified by PCR using human telomerase reverse transcriptase as templates. The amplified fragments were inserted into the eukaryotic expressing vector pcDNA3.1-HisA to construct recombinant plasmid pcDNA3.1-HisA-hTERT aa936-1132. This recombinant plasmid was transfected into the human embryonic kidney 293T cells, and then the protein expression was examined by western blot.
RESULTS AND CONCLUSION: Double enzyme digestion and gene sequencing results showed that the recombinant plasmid pcDNA3.1-HisA-hTERT aa936-1132 containing the C-terminal amino acids 936-1132 of human telomerase reverse transcriptase was constructed successfully. The fusion protein was identified using western blot.