转基因大动物常用载体频数分析及筛查策略

发布时间：2019-06-06 08:16

【摘要】：构建重组表达载体是转基因动物生产制备研究中非常关键的一步,包括构建完整的外源基因表达盒,包含目的基因、调控序列(启动子、终止子)和筛选报告基因等。本文概述了转基因大动物制备技术,归纳统计了近10年转基因猪、牛、羊制备过程中常用的载体和频数,统计结果表明,转基因猪中启动子频数从多到少依次为酪蛋白、CAG、CMV启动子,终止子频数依次为兔β-globin poly A、酪蛋白poly A、SV40poly A和BGH poly A;转基因羊中启动子频数从多到少依次为酪蛋白、BLG和CMV启动子,终止子依次为酪蛋白poly A、BLG poly A、BGH poly A、SV40poly A和兔β-globin poly A;转基因牛中启动子频数从多到少依次为酪蛋白、CMV、人乳清白蛋白启动子等,终止子依次为SV40poly A、BGH poly A和酪蛋白poly A。根据统计结果提出针对启动子和终止子的多重检测和筛查策略;分析了未来转基因动物检测可能存在的问题,以期对转基因动物的监管和筛查检测方法的建立提供参考。
[Abstract]:The construction of recombinant expression vector is a key step in the production and preparation of transgenic animals, including the construction of a complete foreign gene expression cassette, including the target gene, regulatory sequence (promoter, Terminator) and screening reporter gene. In this paper, the preparation techniques of transgenic large animals were summarized, and the vectors and frequencies commonly used in the preparation of transgenic pigs, cattle and sheep in recent 10 years were summarized. The statistical results showed that the promoter frequency of transgenic pigs from more to less was casein, CAG,. The frequency of CMV promoter and Terminator was rabbit 尾-globin poly A, casein poly A, SV40 Poly A and BGH poly A; in turn. The promoter frequency of transgenic sheep was casein, BLG and CMV promoter, and the Terminator was casein poly A, BLG poly A, BGH poly A, SV40 Poly A and rabbit 尾-globin poly A;. The promoter frequency of transgenic cattle was casein, CMV, human milk albumin promoter and so on, and the Terminator was SV40poly A, BGH poly A and casein poly A. According to the statistical results, the multiple detection and screening strategies for promoter and Terminator were put forward, and the possible problems in the detection of transgenic animals in the future were analyzed in order to provide reference for the supervision of transgenic animals and the establishment of screening and detection methods.
【作者单位】：中国检验检疫科学研究院动物检疫研究所;
【基金】：国家科技重大专项(2014ZX0801203B)
【分类号】：S814.8

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