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[Construction of an expression vector under the control of hTERT promoter].

Mingkun Han, Huizhong Zhang, Shi-Yin Cheng

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Abstract

AIM: To construct a luciferase expression vector driven by hTERT(human telomerase reverse transcriptase) core promoter and identify the transcriptional activity of the vector in tumor cells and normal cells. METHODS: hTERT gene core promoter was amplified by PCR using the total genomic DNA from HeLa cells as template. The amplified gene fragment was subsequently cloned into PGL3-basic vector. Then the expression vector was confirmed by restriction enzyme digestion and PCR analysis. The luciferase activity driven by the hTERT gene core promoter was identified after transient transfection of the expression vector into tumor cells and normal cells. RESULTS: The luciferase activity was high in the transfected tumor cells, and very low in transfected normal cells. CONCLUSION: hTERT gene core promoter is tumor-specific and may be useful in gene therapy of tumor.

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What this paper is about

AIM: To construct a luciferase expression vector driven by hTERT(human telomerase reverse transcriptase) core promoter and identify the transcriptional activity of the vector in tumor cells and normal cells. METHODS: hTERT gene core promoter was amplified by PCR using the total genomic DNA from HeLa cells as template. The amplified gene fragment was subsequently cloned into PGL3-basic vector. Then the expression vector was confirmed by restriction enzyme digestion and PCR analysis. The luciferase activity driven by the hTERT gene core promoter was identified after transient transfection of the expression vector into tumor cells and normal cells. RESULTS: The luciferase activity was high in the transfected tumor cells, and very low in transfected normal cells. CONCLUSION: hTERT gene core promoter is tumor-specific and may be useful in gene therapy of tumor.

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Available abstract

AIM: To construct a luciferase expression vector driven by hTERT(human telomerase reverse transcriptase) core promoter and identify the transcriptional activity of the vector in tumor cells and normal cells. METHODS: hTERT gene core promoter was amplified by PCR using the total genomic DNA from HeLa cells as template. The amplified gene fragment was subsequently cloned into PGL3-basic vector. Then the expression vector was confirmed by restriction enzyme digestion and PCR analysis. The luciferase activity driven by the hTERT gene core promoter was identified after transient transfection of the expression vector into tumor cells and normal cells. RESULTS: The luciferase activity was high in the transfected tumor cells, and very low in transfected normal cells. CONCLUSION: hTERT gene core promoter is tumor-specific and may be useful in gene therapy of tumor.

Key concepts: Telomerase reverse transcriptase, Transfection, Luciferase, Molecular biology, Promoter, Vector (molecular biology), Gene, Expression vector

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