陕北白绒山羊转基因试验羊群组织管理与试验辅助工作实施
发布时间：2019-06-22 15:20
【摘要】：为提高陕北白绒山羊绒、肉生产性能,在国家转基因生物新品种培育重大专项支持下,开展了陕北白绒山羊CRISPR/Cas9技术和转基因体细胞核移植技术为技术支撑的转基因试验研究工作。主要针对陕北白绒山羊绒质、肉质性状进行基因改造,以得到遗传性状稳定的优质绒用、肉用陕北白绒山羊品系群,以及优良品系群快速扩繁。在转基因陕北白绒山羊试验研究工作的项目组织中,除上述关键技术环节之外,需要组织规模较大的试验羊群,而试验羊群的准备及饲养管理,转基因受精卵供体母羊超数排卵、配种、胚胎回收,转基因胚胎移植受体同期发情与胚胎移植,转基因胚胎移植受体母羊妊娠期管理、助产乃至剖腹产、产羔母羊和羔羊的围产期管理,以及后代羔羊的哺乳期管理、羔羊培育、生长发育监测和性状监测等工作,是整个转基因项目组织实施中比较重要的环节。这些环节的工作与实验室工作相辅相成,做好这些环节的工作,对提高整个项目实施效率有非常重要的影响,是保证转基因试验项目顺利进行的前提与保障。在陕北白绒山羊转基因试验准备阶段,对试验所需羊群进行组织管理,开展了包括编号、防疫、驱虫、体况评估、供体母羊挑选以及试验羊群发情观察等工作。通过对试验羊群进行发情观察,可知试验羊群发情周期为16~22天,平均为19~20天;试验羊群发情持续期为24~48小时。根据发情周期,选择处于发情周期第4~6天(开始发情记为第0天)的供体母羊作为超数排卵处理对象,进行CIDR预处理,在放置CIDR后第10天7:00第一次肌肉注射Folltropin-V 3.5 mL,19:00第二次肌注Folltropin-V 2.5 mL,第11天7:00第三次肌注Folltropin-V 2.0 mL,19:00第四次Folltropin-V 2.0 mL,第12天7:00第五次肌注Folltropin-V1.5 mL,19:00第六次肌注Folltropin-V 1.0 mL,PG0.1m(1.0mL)g并撤栓。82只超排供体母羊全部进入配种程序,发情正常并配种79只,共回收936枚胚胎,其中可用胚862枚,经CRISPR/Cas9技术进行基因编辑后,416枚胚胎分裂,分裂率为48.25%。在发情周期第4~6天对受体母羊进行NRID预处理,在发情周期第15~17天上午7:00、下午19:00肌注FSH,根据FSH注射剂量不同,将处理受体羊分为三组:第一组76只:FSH33单位,第二组72只:FSH50单位,第三组63只:FSH75单位,发情周期第18天上午7:00撤栓并肌注PG0.1mg。结果显示,不同剂量FSH组同期发情率差异不显著(P0.05);FSH50单位组排卵数较其他两组高(P0.05)。经接产与剖腹产,得到CRISPR/Cas9基因编辑试验羊羔93只。通过定期(每10天一次)测定羊羔体重体尺,得到羊羔生长发育数据,对比前60天转基因阳性羊羔与阴性羊羔生长发育数据,可知,转基因阳性羊羔生长发育快于阴性羊羔。试验工作共获得转基因核移植核移植卵裂胚胎3461枚,移植183只受体。其中58只受体未返情,截至论文成稿共有7只受体流产8只胎儿,足月分娩或剖腹产受体24只,产羔25只。
[Abstract]:In order to improve the performance of white cashmere and meat production in northern Shaanxi, with the support of a major special project for the cultivation of new varieties of transgenic organisms, the transgenic experiment supported by CRISPR/Cas9 technology and transgenic somatic cell nuclear transfer technology in northern Shaanxi white cashmere goat was carried out. Genetic modification of cashmere and meat quality traits of white cashmere goats in northern Shaanxi was carried out in order to obtain high quality cashmere with stable genetic traits, meat white cashmere goat breeds and rapid propagation of fine breeds. In the project organization of transgenic Shaanxi white cashmere goat, in addition to the above key technical links, it is necessary to organize large-scale experimental sheep, and the preparation and feeding management of the experimental sheep, superovulatory, breeding, embryo recovery, simultaneous estrus and embryo transfer of transgenic embryo transfer recipients, pregnancy management, midwifery and even caesarean section of transgenic embryo transfer recipients, The management of parturient period of lambing ewes and lambs, as well as the lactation management of offspring lambs, lamb cultivation, growth and development monitoring and character monitoring, are important links in the implementation of the whole transgenic project organization. The work of these links and the work of the laboratory complement each other. Doing a good job of these links has a very important impact on improving the implementation efficiency of the whole project, and is the premise and guarantee to ensure the smooth progress of the transgenic test project. In the preparation stage of transgenic experiment of white cashmere goat in northern Shaanxi, the sheep needed for the experiment were organized and managed, including numbering, epidemic prevention, insect repellent, body condition evaluation, selection of donor ewes and estrus observation of experimental sheep. Through the estrus observation of the experimental sheep, it was found that the estrus cycle of the experimental sheep was 16 脳 22 days, with an average of 19 脳 20 days, and the estrus duration of the experimental sheep was 24 鈮，
本文编号：2504705
[Abstract]:In order to improve the performance of white cashmere and meat production in northern Shaanxi, with the support of a major special project for the cultivation of new varieties of transgenic organisms, the transgenic experiment supported by CRISPR/Cas9 technology and transgenic somatic cell nuclear transfer technology in northern Shaanxi white cashmere goat was carried out. Genetic modification of cashmere and meat quality traits of white cashmere goats in northern Shaanxi was carried out in order to obtain high quality cashmere with stable genetic traits, meat white cashmere goat breeds and rapid propagation of fine breeds. In the project organization of transgenic Shaanxi white cashmere goat, in addition to the above key technical links, it is necessary to organize large-scale experimental sheep, and the preparation and feeding management of the experimental sheep, superovulatory, breeding, embryo recovery, simultaneous estrus and embryo transfer of transgenic embryo transfer recipients, pregnancy management, midwifery and even caesarean section of transgenic embryo transfer recipients, The management of parturient period of lambing ewes and lambs, as well as the lactation management of offspring lambs, lamb cultivation, growth and development monitoring and character monitoring, are important links in the implementation of the whole transgenic project organization. The work of these links and the work of the laboratory complement each other. Doing a good job of these links has a very important impact on improving the implementation efficiency of the whole project, and is the premise and guarantee to ensure the smooth progress of the transgenic test project. In the preparation stage of transgenic experiment of white cashmere goat in northern Shaanxi, the sheep needed for the experiment were organized and managed, including numbering, epidemic prevention, insect repellent, body condition evaluation, selection of donor ewes and estrus observation of experimental sheep. Through the estrus observation of the experimental sheep, it was found that the estrus cycle of the experimental sheep was 16 脳 22 days, with an average of 19 脳 20 days, and the estrus duration of the experimental sheep was 24 鈮，
本文编号：2504705
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