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Cre介导的斑马鱼胚胎位点特异性重组。

Cre-mediated site-specific recombination in zebrafish embryos.

作者信息

Thummel Ryan, Burket Christopher T, Brewer Jeffrey L, Sarras Michael P, Li Li, Perry Martin, McDermott Jeffrey P, Sauer Brian, Hyde David R, Godwin Alan R

机构信息

Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, USA.

出版信息

Dev Dyn. 2005 Aug;233(4):1366-77. doi: 10.1002/dvdy.20475.

Abstract

Cre-mediated site-specific recombination has become an invaluable tool for manipulation of the murine genome. The ability to conditionally activate gene expression or to generate chromosomal alterations with this same tool would greatly enhance zebrafish genetics. This study demonstrates that the HSP70 promoter can be used to inducibly control expression of an enhanced green fluorescent protein (EGFP) -Cre fusion protein. The EGFP-Cre fusion protein is capable of promoting recombination between lox sites in injected plasmids or in stably inherited transgenes as early as 2 hr post-heat shock induction. Finally, the levels of Cre expression achieved in a transgenic fish line carrying the HSP70-EGFP-cre transgene are compatible with viability and both male and female transgenic fish are fertile subsequent to induction of EGFP-Cre expression. Hence, our data suggests that Cre-mediated recombination is a viable means of manipulating gene expression in zebrafish.

摘要

Cre介导的位点特异性重组已成为操纵小鼠基因组的一项极有价值的工具。利用这一相同工具来条件性激活基因表达或产生染色体改变的能力将极大地增强斑马鱼遗传学研究。本研究表明,热休克蛋白70(HSP70)启动子可用于诱导性控制增强型绿色荧光蛋白(EGFP)-Cre融合蛋白的表达。早在热休克诱导后2小时,EGFP-Cre融合蛋白就能促进注射质粒或稳定遗传转基因中lox位点之间的重组。最后,携带HSP70-EGFP-cre转基因的转基因鱼系中实现的Cre表达水平与生存能力相匹配,并且在诱导EGFP-Cre表达后,转基因雄鱼和雌鱼均具有生育能力。因此,我们的数据表明,Cre介导的重组是在斑马鱼中操纵基因表达的一种可行方法。

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