中国科学院水生生物研究所机构知识库
Advanced  
IHB OpenIR  > 中科院水生所知识产出(2009年前)  > 期刊论文
学科主题: Biotechnology & Applied Microbiology; Marine & Freshwater Biology
题名: RecA-mediated, targeted mutagenesis in zebrafish
作者: Cui, ZB; Yang, Y; Kaufman, CD; Agalliu, D; Hackett, PB
通讯作者: Yang, Y, Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA
关键词: RecA protein ; gene targeting ; site-specific mutagenesis ; zebrafish
刊名: MARINE BIOTECHNOLOGY
发表日期: 2003-03-01
DOI: 10.1007/s10126-002-0059-0
卷: 5, 期:2, 页:174-184
收录类别: SCI
文章类型: Article
部门归属: Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA; Univ Minnesota, Arnold & Mabel Beckman Ctr Transposon Res, St Paul, MN 55108 USA; Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
研究领域[WOS]: Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
摘要: We have evaluated the efficacy of RecA, a prokaryotic protein involved with homologous recombination, to direct site-specific mutagenesis in zebrafish embryos. For this we coinjected a vector containing a mutated enhanced green fluorescent protein (EGFP) gene plus 236-nucleotide corrective single-stranded DNAs coated with RecA into I-cell zebrafish embryos. Twenty-hours after fertilization, about 5% to 20% of injected embryos showed EGFP expression in I or more cells when RecA-coated corrective DNAs were used, but not when RecA was omitted. Mutated EGFP genes with 1-bp insertions or deletions were inefficiently activated, whereas those with 7-bp insertions were activated about 4-fold more efficiently. RecA-coated template strand had a higher efficiency than its complementary strand in activation of EGFP expression. Prior irradiation of the embryos with UV light enhanced RecA-mediated restoration of gene activity, suggesting that the effects we observed were augmented by one or more factors of zebrafish DNA repair systems.
英文摘要: We have evaluated the efficacy of RecA, a prokaryotic protein involved with homologous recombination, to direct site-specific mutagenesis in zebrafish embryos. For this we coinjected a vector containing a mutated enhanced green fluorescent protein (EGFP) gene plus 236-nucleotide corrective single-stranded DNAs coated with RecA into I-cell zebrafish embryos. Twenty-hours after fertilization, about 5% to 20% of injected embryos showed EGFP expression in I or more cells when RecA-coated corrective DNAs were used, but not when RecA was omitted. Mutated EGFP genes with 1-bp insertions or deletions were inefficiently activated, whereas those with 7-bp insertions were activated about 4-fold more efficiently. RecA-coated template strand had a higher efficiency than its complementary strand in activation of EGFP expression. Prior irradiation of the embryos with UV light enhanced RecA-mediated restoration of gene activity, suggesting that the effects we observed were augmented by one or more factors of zebrafish DNA repair systems.
关键词[WOS]: STIMULATES HOMOLOGOUS RECOMBINATION ; DNA STRAND-EXCHANGE ; TRANSGENIC ZEBRAFISH ; ESCHERICHIA-COLI ; ORYZIAS-LATIPES ; MAMMALIAN-CELLS ; PROTEIN ; REPAIR ; GENE ; EXPRESSION
语种: 英语
WOS记录号: WOS:000182588700011
ISSN号: 1436-2228
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/9700
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

Files in This Item: Download All
File Name/ File Size Content Type Version Access License
RecA-mediated, targeted mutagenesis in zebrafish.pdf(260KB)----开放获取--View Download

作者单位: 1.Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA
2.Univ Minnesota, Arnold & Mabel Beckman Ctr Transposon Res, St Paul, MN 55108 USA
3.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China

Recommended Citation:
Zongbin Cui; Ying Yang; Christopher D. Kaufman; Dritan Agalliu; Perry B. Hackett.RecA-mediated, targeted mutagenesis in zebrafish,MARINE BIOTECHNOLOGY,2003,5(2):174-184
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[Cui, ZB]'s Articles
[Yang, Y]'s Articles
[Kaufman, CD]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[Cui, ZB]‘s Articles
[Yang, Y]‘s Articles
[Kaufman, CD]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
文件名: RecA-mediated, targeted mutagenesis in zebrafish.pdf
格式: Adobe PDF
此文件暂不支持浏览
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Copyright © 2007-2017  中国科学院水生生物研究所 - Feedback
Powered by CSpace