文章摘要
胡莹莹,郭学双,周 阳,曹广力,贡成良,薛仁宇.piggyBac转座子在金鱼及泥鳅转基因中的应用[J].水产学报,2012,36(10):1473~1481
piggyBac转座子在金鱼及泥鳅转基因中的应用
Exploration of transgenic method for goldfish and loach with the piggyBac transposon
投稿时间:2012-01-05  修订日期:2012-06-27
DOI:10.3724/SP.J.1231.2012.27930
中文关键词: 金鱼  泥鳅  piggyBac转座子  转基因  草鱼肾脏(CIK)
英文关键词: piggyBac transposon  transgene  Ctenpharyngoden Idellus kidney, CIK  Carassius auratus  Misgurnus anguillicaudatus
基金项目:国家重大基础研究项目(2012CB114605)
作者单位
胡莹莹 苏州大学基础医学与生物科学学院 
郭学双 苏州大学基础医学与生物科学学院 
周 阳 苏州大学基础医学与生物科学学院 
曹广力 苏州大学基础医学与生物科学学院 
贡成良 苏州大学基础医学与生物科学学院 
薛仁宇 苏州大学基础医学与生物科学学院 
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中文摘要:
      为了探讨piggyBac转座子在鲤科鱼类及鳅科鱼类转基因中的适用性,实验通过将PAβ启动子控制的DsRed表达元件插入pigA3GFP中,构建转基因载体pigA3GFP AβDsRed。用该载体对草鱼肾脏(CIK)细胞转染,结果显示,通过piggyBac转座子可将外源基因导入到CIK细胞的基因组中,来源于家蚕的A3启动子和来源于巨细胞病毒的CMV启动子在CIK细胞均具有活性。将转基因载体pigA3GFP、pigA3GFP AβDsRed分别与辅助质粒helper pigA3混合后以精子介导法导入金鱼和泥鳅的受精卵后,经荧光观察、PCR鉴定、Dot blotting鉴定,证实获得了转基因金鱼和转基因泥鳅。研究表明,PB转座子在鲤科鱼类及鳅科的一些鱼中具有转座活性。
英文摘要:
      To investigate the applicability of the transposon piggyBac in Cyprinidae and Cobitidae fish, the expression element of DsRed under the control of PAβ promoter was inserted into the plasmid pigA3GFP to generate transgenic vector pigA3GFP-AβDsRed. The results of the CIK cell transfected with pigA3GFP-AβDsRed showed that the transposon piggyBac could transfer exogenous genes into CIK genome, and both of the promoter A3 from silkworm and the promoter CMV from CMV have the bioactivity. Moreover, the pigA3GFP-AβDsRed and pigA3GFP were mixed with helper plasmid helper-pigA3 respectively, and then were introduced into the eggs of goldfish and loach by sperm-mediated transfer. The results of fluorescence observation, PCR and Dot blotting verified that both transgenic goldfish and loach were obtained. Therefore, PB transposon was considered to be active in some fish of Cyprinidae and Cobitidae. Therefore, PB transposon was considered to be active in some fish of Cyprinidae and Cobitidae.
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