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Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4

机译：在繁殖和非繁殖季节高效生产原核胚胎，以产生过表达TLR4的转基因绵羊

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摘要

Background:Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans.TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity.Consequently,transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis.In this study,we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons (breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.Results:In total of 43 donor ewes and 166 recipient ewes in breeding season,37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no.of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons (P ＞ 0.05).The positive rate of exogenous TLR4 tested were 21.21％ and 22.58 ％ in breeding and non-breeding season by Southern blot.The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group (P ＜ 0.05).The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs,and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions:Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year The over-expression of TLR4 had no adverse effect on the growth of the sheep.
机译：背景：布鲁氏菌是一种人畜共患的革兰氏阴性病原体，可导致反刍动物和人类流产和不育.TLR4是LPS的受体，可以识别布鲁氏菌并启动影响天然免疫和适应性免疫的抗原呈递细胞活性。表达TLR4是研究TLR4预防布鲁氏菌病的合适模型。在本研究中，我们产生了过表达TLR4的转基因绵羊，旨在评估不同季节（繁殖和非繁殖季节）对超排卵和外源外源输入的影响。结果：繁殖季节共有43只供体母羊和166只受体母羊，非繁殖季节选择了37只供体母羊和144只受体母羊进行超排卵，并进行胚胎移植以产生转基因绵羊。捐献者的胚胎数和原核胚胎的发生率之间没有显着差异。 d非繁殖季节（P＞ 0.05）。通过Southern blot检测，在繁殖和非繁殖季节外源TLR4的阳性率分别为21.21％和22.58％。转基因绵羊中TLR4的表达水平是转基因绵羊的1.5倍。非转基因组（P ＜0.05）。过表达TLR4的羔羊的生长性能与非转基因羔羊相似，转基因和非转基因的血液生理指标均在正常范围内，无任何差异。 ：在这里，我们建立了一个全年通过显微注射原核胚胎生产转基因绵羊的有效平台。TLR4的过表达对绵羊的生长没有不利影响。

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  • 来源
    《畜牧与生物技术杂志(英文版)》 |2017年第1期|43-50|共8页
  • 作者

    Yan Li; Juncai Fu; Hongbing Han; Jianzhong Feng; Guoshi Liu; Ling Lian; Zhengxing Lian; Di Lian; Shoulong Deng; Xiaosheng Zhang; Jinlong Zhang; Wenting Li; Hai Bai; Zhixian Wang; Hongping Wu;

  • 作者单位

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Tianjin Institute of Animal Sciences, Tianjin 300381, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Department of Public Health, Benedictine University, Lisle, IL 60532, USA;

    State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China;

    Tianjin Institute of Animal Sciences, Tianjin 300381, China;

    Tianjin Institute of Animal Sciences, Tianjin 300381, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;

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