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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.

RESULTS AND CONCLUSION:

Lu JX, Jiang HX, Cao Y, Pang JX. Construction of an eukaryotic expression vector carrying amino acids 936-1132 of human telomerase reverse transcriptase and its expression in 293T cells. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(11): 1977-1980. [http://www.crter.cnhttp://en.zglckf.com]

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