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转生长激素基因中华倒刺鲃的构建及其检测
引用本文:丁洁,顾继锐,伍翠芳,李永文,张涛,吴江,谢显连,徐恒.转生长激素基因中华倒刺鲃的构建及其检测[J].淡水渔业,2006,36(3):30-33.
作者姓名:丁洁  顾继锐  伍翠芳  李永文  张涛  吴江  谢显连  徐恒
作者单位:生物资源与生态环境教育部重点实验室,四川大学生命科学学院,成都,610064;成都通威水产科技有限公司基因实验室,成都,610081;成都通威水产科技有限公司基因实验室,成都,610081;生物资源与生态环境教育部重点实验室,四川大学生命科学学院,成都,610064
基金项目:水利部“948”项目,通威股份科技发展项目
摘    要:用从斑点叉尾鮰(Ictalurus punctatus)cDNA文库克隆到的生长激素基因,与鲤β肌蛋白启动子、大麻哈鱼生长激素基因的poly(A)终止信号序列等调控元件共同构建了表达重组质粒pFV2 cfGH。提取重组质粒,用精子载体法导入中华倒刺鲃(Spinibarbus sinensis)的受精卵中,经孵化培养成转基因成鱼。抽提423尾转基因实验鱼血总DNA,经PCR检测表明:斑点叉尾鮰生长激素基因已导入中华倒刺鲃基因组中,导入率为17.26%。同时通过RT-PCR技术检测阳性中华倒刺鲃中生长激素基因的表达情况。

关 键 词:生长激素基因  中华倒刺鲃  转基因鱼  反转录PCR
文章编号:1000-6907-(2006)03-0030-04
收稿时间:2006-03-06
修稿时间:2006年3月6日

The Construction and Detecting of Transgenic Spinibarbus sinensis With Growth Hormone Gene
DING Jie,GU Ji-rui,WU Cui-fang,LI Yong-wen,ZHANG Tao,WU Jiang,XIE Xian-lian,XU Heng.The Construction and Detecting of Transgenic Spinibarbus sinensis With Growth Hormone Gene[J].Freshwater Fisheries,2006,36(3):30-33.
Authors:DING Jie  GU Ji-rui  WU Cui-fang  LI Yong-wen  ZHANG Tao  WU Jiang  XIE Xian-lian  XU Heng
Institution:1. Key laboratory of Bio - resources and Eco - environment( Ministry of Education
Abstract:The expression recombinant plasmid pFV2cfGH was constructed from growth hormone gene which was cloned from the channel catfish cDNA library,the β-action promoter of carp and the polyadenylation signal from the salmon growth hormone gene,and other controlling elements.The recombinant plasmid pFV2cfGH was transmitted into the fertilized eggs of the Spinibarbus sinensis through Sperm-mediated gene transfer,and was hatched into transgenic fish.Total DNA of 423 transgenic Spinibarbus sinensis was abstracted,and growth hormone gene was detected by PCR.The results showed that the growth hormone gene of the channelfish had been transmitted into the genome of the Spinibarbus sinensis and the integration rate is 17.26%.At the same time,the expression of growth hormone gene in positive Spinibarbus sinensis was detected by RT-RCR.
Keywords:growth hormone gene  Spinibarbus sinensis  transgenic fish  RT-PCR
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