Functional characterization of the zebrafish gadd45αb gene promoter and its application as a biosensor

被引：0
|
作者：
Gireesh-Babu, P. [1 ]
Pawar, Nilambari [1 ]
Krishnan, P. [1 ]
Kumar, A. Pavan [1 ]
Zaidi, S. G. S. [1 ]
Bhartiya, Deepa [2 ]
Kumar, Neeraj [2 ]
Sivasubbu, Sridhar [3 ]
Rajendran, K. V. [1 ]
Chaudhari, Aparna [1 ]
机构：
[1] ICAR Res Complex, Cent Inst Fisheries Educ, Bombay 400061, Maharashtra, India
[2] ICMR, Natl Inst Res Reprod Hlth, Bombay 400012, Maharashtra, India
[3] CSIR, Inst Genom & Integrat Biol, New Delhi 110007, India
来源：
CURRENT SCIENCE | 2012年 / 103卷 / 04期
关键词：
Biosensor; gadd45; alpha; genotoxicity; minimal promoter; zebrafish; DNA; KINASE; GFP; INDUCTION; CADMIUM; DAMAGE; OCT-1; ASSAY;
D O I：
暂无
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
We report characterization of zebrafish gadd45 alpha b (growth arrest and DNA damage gene) minimal promoter and its application as biosensor tool for genotoxicity studies in zebrafish. Variably sized fragments of the 5' flanking region were generated using polymerase chain reaction (PCR). These were cloned upstream of gfp gene in pEGFP1 vector, transfected into human foetal fibroblasts and induced with DNA damaging agents, ultraviolet radiation (UV) and methyl methane sulphonate (MMS). The region -1064/+593 bp showed maximum activity after treatment with 1.5 mM MMS for 2 h or 80 mJ/cm(2) UV. The endogenous gadd45 alpha expression in human fibroblast cells and zebrafish embryos in the presence of UV and MMS was confirmed by real-time PCR. When the minimal promoter linked to dsRed2 gene was microinjected into zebrafish embryos, red fluorescence was observed in response to sub-lethal dose of UV, confirming its application as transgenic zebrafish biosensor for genotoxicity.
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页码：388 / 394
页数：7
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