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Tgf2转座系统在转RFP基因斑马鱼上的应用
作者:
作者单位:

1. 中国水产科学研究院 珠江水产研究所; 农业部热带亚热带水产资源利用与养殖重点实验室, 广东 广州 510380;

作者简介:

吴芳(1988–), 女, 硕士研究生, 从事水产生物技术研究. E-mail: wfone0305@163.com 通信作者: 叶星, 研究员, 博士生导师. E-mail: gzyexing@163.com

中图分类号:

S962.1

基金项目:

国家863计划项目(2011AA100404); 国家自然科学资金面上项目(31272688).


Application of the Tgf2 transposon system to RFP transgenic Danio rerio construction
Author:
Affiliation:

1. Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences of China, Key Laboratory of Tropic & Subtropic Fisheries Resource Utilization Aquaculture, Ministry of Agriculture, Guangzhou 510380, China;

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    摘要:

    提高目的基因的转植效率与可遗传效率是转基因鱼研制的关键点之一。本研究利用近年开发的金鱼转座子系统进行转基因斑马鱼的研制将肌球蛋白轻链肌肉组织特异性表达红色荧光蛋白的转座子介导的转基因技术

    Abstract:

    Improving the efficiency of gene transfer and integration is a key objective in transgenic fish research. To evaluate its use for transgenic fish applications, we used the Tgf2 transposon donor plasmid pTgf2-EF1α-eGFP was re-constructed by replacement of the transgenic elements, EF1α-eGFP with the myosin light chain 2 promoter (mylz2) and the red fluorescent protein (RFP) gene, mylz2-RFP. The recombinant transposon donor plasmid, pTgf2-Mylz2-RFP was intended to serve as a transposon donor plasmid that specifically expresses red fluorescent protein in muscle tissue. The donor plasmid and the transposase mRNA were co-injected into zebra fish fertilized eggs. A total of 972 eggs were microinjected and 803 larvae survived, with a positive rate of 76.6%. The red fluorescence () were subsequently cultivated and 10 sexually mature transgenic individuals were mated with wild-type zebrafish, respectively. F1 individuals that exhibited red fluorescence on their body surface were obtained from one of the mating pairs yielding an integration efficiency of 10%. The positive rate of F2 individuals, which were generated by mating the mature transgenic F1 with the wild-type fish, was 69%. The mature RFP positive F1 individual was then mated with wild-type fish, yielding a 69% positive rate in the F2 generation. Our results demonstrate that the transposon improves the efficiency of gene transfer and integration during transgene treatments. Thus, the transposon can be used to construct transgenic fish and in other transgenic fish research.

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吴芳,叶星,邹曙明,张莉莉,孙成飞,田园园,白俊杰. Tgf2转座系统在转RFP基因斑马鱼上的应用[J].中国水产科学,2014,21(4):647-654
WU Fang, YE Xing, ZOU Shuming, ZHANG Lili, SUN Chengfei, TIAN Yuanyuan, BAI Junjie. Application of the Tgf2 transposon system to RFP transgenic Danio rerio construction[J]. Journal of Fishery Sciences of China,2014,21(4):647-654

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  • 在线发布日期: 2015-08-04
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