陜北白絨山羊轉(zhuǎn)基因試驗(yàn)羊群組織管理與試驗(yàn)輔助工作實(shí)施
發(fā)布時(shí)間：2019-06-22 15:20
【摘要】：為提高陜北白絨山羊絨、肉生產(chǎn)性能,在國家轉(zhuǎn)基因生物新品種培育重大專項(xiàng)支持下,開展了陜北白絨山羊CRISPR/Cas9技術(shù)和轉(zhuǎn)基因體細(xì)胞核移植技術(shù)為技術(shù)支撐的轉(zhuǎn)基因試驗(yàn)研究工作。主要針對陜北白絨山羊絨質(zhì)、肉質(zhì)性狀進(jìn)行基因改造,以得到遺傳性狀穩(wěn)定的優(yōu)質(zhì)絨用、肉用陜北白絨山羊品系群,以及優(yōu)良品系群快速擴(kuò)繁。在轉(zhuǎn)基因陜北白絨山羊試驗(yàn)研究工作的項(xiàng)目組織中,除上述關(guān)鍵技術(shù)環(huán)節(jié)之外,需要組織規(guī)模較大的試驗(yàn)羊群,而試驗(yàn)羊群的準(zhǔn)備及飼養(yǎng)管理,轉(zhuǎn)基因受精卵供體母羊超數(shù)排卵、配種、胚胎回收,轉(zhuǎn)基因胚胎移植受體同期發(fā)情與胚胎移植,轉(zhuǎn)基因胚胎移植受體母羊妊娠期管理、助產(chǎn)乃至剖腹產(chǎn)、產(chǎn)羔母羊和羔羊的圍產(chǎn)期管理,以及后代羔羊的哺乳期管理、羔羊培育、生長發(fā)育監(jiān)測和性狀監(jiān)測等工作,是整個(gè)轉(zhuǎn)基因項(xiàng)目組織實(shí)施中比較重要的環(huán)節(jié)。這些環(huán)節(jié)的工作與實(shí)驗(yàn)室工作相輔相成,做好這些環(huán)節(jié)的工作,對提高整個(gè)項(xiàng)目實(shí)施效率有非常重要的影響,是保證轉(zhuǎn)基因試驗(yàn)項(xiàng)目順利進(jìn)行的前提與保障。在陜北白絨山羊轉(zhuǎn)基因試驗(yàn)準(zhǔn)備階段,對試驗(yàn)所需羊群進(jìn)行組織管理,開展了包括編號、防疫、驅(qū)蟲、體況評估、供體母羊挑選以及試驗(yàn)羊群發(fā)情觀察等工作。通過對試驗(yàn)羊群進(jìn)行發(fā)情觀察,可知試驗(yàn)羊群發(fā)情周期為16~22天,平均為19~20天;試驗(yàn)羊群發(fā)情持續(xù)期為24~48小時(shí)。根據(jù)發(fā)情周期,選擇處于發(fā)情周期第4~6天(開始發(fā)情記為第0天)的供體母羊作為超數(shù)排卵處理對象,進(jìn)行CIDR預(yù)處理,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只超排供體母羊全部進(jìn)入配種程序,發(fā)情正常并配種79只,共回收936枚胚胎,其中可用胚862枚,經(jīng)CRISPR/Cas9技術(shù)進(jìn)行基因編輯后,416枚胚胎分裂,分裂率為48.25%。在發(fā)情周期第4~6天對受體母羊進(jìn)行NRID預(yù)處理,在發(fā)情周期第15~17天上午7:00、下午19:00肌注FSH,根據(jù)FSH注射劑量不同,將處理受體羊分為三組:第一組76只:FSH33單位,第二組72只:FSH50單位,第三組63只:FSH75單位,發(fā)情周期第18天上午7:00撤栓并肌注PG0.1mg。結(jié)果顯示,不同劑量FSH組同期發(fā)情率差異不顯著(P0.05);FSH50單位組排卵數(shù)較其他兩組高(P0.05)。經(jīng)接產(chǎn)與剖腹產(chǎn),得到CRISPR/Cas9基因編輯試驗(yàn)羊羔93只。通過定期(每10天一次)測定羊羔體重體尺,得到羊羔生長發(fā)育數(shù)據(jù),對比前60天轉(zhuǎn)基因陽性羊羔與陰性羊羔生長發(fā)育數(shù)據(jù),可知,轉(zhuǎn)基因陽性羊羔生長發(fā)育快于陰性羊羔。試驗(yàn)工作共獲得轉(zhuǎn)基因核移植核移植卵裂胚胎3461枚,移植183只受體。其中58只受體未返情,截至論文成稿共有7只受體流產(chǎn)8只胎兒,足月分娩或剖腹產(chǎn)受體24只,產(chǎn)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，
本文編號：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，
本文編號：2504705
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