Isolation and Activity Analysis of a Minimal Promoter of the Human Tyrosine Hydroxylase Gene

doi:10.3969/j.issn.1006-7795.2009.05.005

Journal of Capital Medical University ›› 2009, Vol. 30 ›› Issue (5): 593-596.doi: 10.3969/j.issn.1006-7795.2009.05.005

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Isolation and Activity Analysis of a Minimal Promoter of the Human Tyrosine Hydroxylase Gene

LI Yao-hua¹, YE Yi-wen¹, GAO Nan¹, LI Xin¹, YU Shun¹, YANG Hui², CHEN Biao¹

1. Department of Neurobiology, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Diseases of Ministry of Education of China;2. Institute for Neuroscience, Capital Medical University

Received:2009-07-16 Revised:1900-01-01 Online:2009-10-21 Published:2009-10-21

Abstract

Abstract: Objective To investigate the regulatory functions of human tyrosine hydroxylase gene upstream region. Methods A human TH gene upstream fragment was amplified by PCR, and was inserted into plasmid pGL3-Basic, to build a luciferase reporter vector. The reporter vector was transient transfected into MES23.5, 293T and Cos7 cells, and the promoter activity was mensurated using Dual Luciferase Assay system. Results Length of the isolated DNA fragment is 520 bp, which located at -495/+25 region of the human tyrosine hydroxylase gene. By comparison with pGL3-Basic vector, the recombinant reporter vector could significantly expressed luciferase in three types of cells(P<0.01). Conclusion TH gene -493/+27 region possesses promoter activity, and the constructed reporter gene will help us study the regulation of TH gene expression.

Key words: tyrosine hydroxylase, promoter, dopamine, Parkinson's disease

CLC Number:

Q 71

LI Yao-hua;YE Yi-wen;GAO Nan;LI Xin;YU Shun;YANG Hui;CHEN Biao. Isolation and Activity Analysis of a Minimal Promoter of the Human Tyrosine Hydroxylase Gene[J]. Journal of Capital Medical University, 2009, 30(5): 593-596.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2009.05.005

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2009/V30/I5/593

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Isolation and Activity Analysis of a Minimal Promoter of the Human Tyrosine Hydroxylase Gene

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