摘要
[目的]研究不同的胚龄、移植季节以及胚胎数量对转人乳铁蛋白基因（hLF）体细胞克隆山羊生产效率的影响.[方法]用奶山羊的转hLF基因成纤维细胞为供体细胞生产克隆胚胎,并以不同的胚龄、胚胎数量,在不同的季节移植给同期的受体山羊.[结果]移植1细胞期和2-8细胞期胚胎受体羊的妊娠率（分别是17.95%和28.57%）、产羔率（分别是12.82%和14.29%）显著高于移植桑椹胚、囊胚的受体羊（3.70%和0,P〈0.05）;转hLF基因克隆胚胎在秋季移植效率最高,虽然秋季和春季受体羊妊娠率（分别是19.57%和18.18%）与冬季（10.34%）差异不显著,但秋季受体羊产羔率（13.04%）显著高于冬季（1.72%,P〈0.05）;虽然移植10-20枚克隆胚胎的受体羊妊娠率（18.18%）显著高于移植5-9枚胚胎的受体羊（7.89%）,但是由于其流产率高,两者产羔率差异并不显著（7.89% vs 6.36%,P〉0.05）.[结论]转基因克隆山羊胚胎移植试验的最适季节是秋季,最好移植激活后体外培养2 d内的胚胎.
[Objectives]Embryo transfer is one of the important factors that influence the efficiency of transgenic cloned goat production and three factors affect the embryo transfer：the embryo age,season and number of embryo transferred. The aim of the paper is to investigate the effect of the embryo age,season and number of embryo transferred on the efficiency of human lactoferrin（hLF）transgenic cloned goat production. [Methods]Cloned embryos using dairy goat hLF transgenic fibroblast cells as donor cells at different stage and number were transferred to the uterus of recipient goat at different seasons. The current study was conducted from October to April next year,lasting for seven months. First,estrus synchronization and superovulation were carried out. Secondly,somatic cell nuclear transfer（SCNT）was performed according to the standard protocol using dairy goat hLF transgenic fibroblast cells as donor cells. Lastly,the constructed embryos were transferred to the uterus of recipient goat. [Results]Our results showed that both the pregnancy rate and the kidding rate of the recipient with one-cell（17.95%,12.82%,respectively）and 2-8 cell embryo（28.57%,14.29%,respectively）transferred were higher than those of the recipient with morale or blastocyst transferred（3.70%,0,respectively,P〈0.05）. Embryo transfer was conducted during spring,autumn and winter,and we found that the efficiency was higher during autumn and the pregnancy rate of the recipient was not affected by the season,while the kidding rate of the recipient was higher in autumn than that in winter（13.04% vs 1.72%,P〈0.05）. Moreover,the pregnancy rate of the recipient with 10-20 cloned embryos transferred was significantly greater than that with 5-9 cloned embryos transferred（18.18% vs 7.89%,P〈0.05）,however,the abortion was also higher in the former group. Thus,there was no significant difference in the kidding rate between these two groups. To test the development ability of constructed embryos,the embryos were coated by agarose and transferred to uterus for a certain time and rushed out. Taken all these together,our experiment indicated that intermediate receptor method can be used to determine the embryo quality and the number of embryos transferred（7.89% vs 6.36%,P〉0.05）. [Conclusions]Our current work revealed that the most suitable season for hLF-transgenic cloned goat production is autumn and the embryos transfer should be conducted within 2 days after the activation. This work might lay a foundation for the research of SCNT efficiency.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2015年第6期980-985,共6页
Journal of Nanjing Agricultural University
基金
国家自然科学基金项目(31301973)
中央高校基本科研业务费专项资金项目(KJQN201401)
国家转基因新品种培育重大专项(2014ZX08008-003
2014ZX08008-004)
关键词
转基因克隆
胚胎移植
妊娠率
产羔率
奶山羊
transgenic clone
embryo transfer
pregnancy rate
kidding rate
dairy goat
作者简介
万永杰，讲师，主要从事动物胚胎工程研究。
通信作者：黄明睿，副编审，主要从事肉羊繁育研究及畜牧兽医技术推广，E—mail：hmr@njau．edu．cn。
