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[Construction of recombinant pEGFP/hTRT plasmid].

Hai-wen Zhou¹, Zeng-tong Zhou, Wen-jun Yang

¹Department of Oral Medicine, Ninth People's Hospital, Shanghai Second Medical University, Shanghai, P. R. China 200011.

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery

|June 26, 2003

Summary

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Researchers successfully created a new plasmid, pEGFP-hTRT, to study human telomerase reverse transcriptase (hTRT) in cell cultures. This tool aids in developing immortalized cell lines.

Area of Science:

Molecular Biology
Cell Biology
Genetics

Context:

Human telomerase reverse transcriptase (hTRT) plays a crucial role in cellular processes, including cell immortalization.
Understanding hTRT's function is vital for research in aging and cancer.
In vitro cell cultivation requires tools to study specific gene functions.

Purpose:

To construct a novel plasmid vector for the expression and study of human telomerase reverse transcriptase (hTRT).
To facilitate the exploration of hTRT's role in in vitro cell cultivation and immortalization.
To develop a reporter gene system for monitoring hTRT activity.

Summary:

The human telomerase reverse transcriptase (hTRT) gene was isolated from plasmid pGRN145 using restriction enzymes.

Related Experiment Videos

The isolated hTRT gene was successfully inserted into the pEGFP-C1 plasmid, creating the pEGFP-hTRT construct.

Restriction enzyme analysis and DNA sequencing confirmed the accurate sequence of the pEGFP-hTRT transgenic plasmid.

Impact:

The successful construction of the pEGFP-hTRT recombinant vector provides a valuable tool for generating immortalized cell lines.
This transgenic plasmid enables further investigation into the mechanisms of hTRT and its implications in cellular aging and disease.
Facilitates advancements in cell-based research and therapeutic development.