体细胞克隆技术扩繁超细型二狼山绒山羊的应用研究

doi:10.16431/j.cnki.1671-7236.2024.04.010

中国畜牧兽医 ›› 2024, Vol. 51 ›› Issue (4): 1420-1427.doi: 10.16431/j.cnki.1671-7236.2024.04.010

• 生物技术 • 上一篇

体细胞克隆技术扩繁超细型二狼山绒山羊的应用研究

卿玉波^1,2, 程雯^1,2, 角德灵^1,2, 刘城^1,3, 郝志强⁴, 郭建雄¹, 李钰莹^1,3, 张筱琳^1,3, 李臻^4,5, 李效宇^4,5, 王文⁶, 魏红江^1,2

1. 云南省小型猪基因编辑与异种器官移植重点实验室, 昆明 650201;
2. 云南农业大学动物医学院, 昆明 650201;
3. 云南农业大学动物科学技术学院, 昆明 650201;
4. 内蒙古中科正标生物科技有限公司, 巴彦淖尔 015000;
5. 内蒙古正标种畜有限公司, 巴彦淖尔 015000;
6. 中国科学院昆明动物研究所, 昆明 650201

收稿日期:2023-09-05 发布日期:2024-03-27
通讯作者: 魏红江 E-mail:hongjiangwei@126.com
作者简介:卿玉波,E-mail:qingyubo20@163.com;程雯,E-mail:chengcheng20200921@163.com;角德灵,E-mail:jiaodeling@163.com。
基金资助:
云南省国际科技合作专项-巴基斯坦肉羊扩繁技术集成、应用与推广(2016IA024)

Study on the Application of Propagation of Superfine-type Erlangshan Cashmere Goat by Somatic Cell Cloning Technology

QING Yubo^1,2, CHENG Wen^1,2, JIAO Deling^1,2, LIU Cheng^1,3, HAO Zhiqiang⁴, GUO Jianxiong¹, LI Yuying^1,3, ZHANG Xiaolin^1,3, LI Zhen^4,5, LI Xiaoyu^4,5, WANG Wen⁶, WEI Hongjiang^1,2

1. Key Laboratory for Porcine Gene Editing and Xenotransplantation in Yunnan Province, Kunming 650201, China;
2. College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, China;
3. Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
4. Inner Mongolia Zhongke Zhengbiao Biotechnology Co., Ltd., Bayannur 015000, China;
5. Inner Mongolia Zhengbiao Breeding Stock Co., Ltd., Bayannur 015000, China;
6. Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650201, China

Received:2023-09-05 Published:2024-03-27

摘要/Abstract

摘要： 【目的】通过体细胞克隆技术生产超细型二狼山绒山羊克隆个体,为该技术在地方优质羊品种资源扩繁及新品种培育中的应用提供理论依据。【方法】选择1只4岁龄羊绒细度为13.89 μm的二狼山绒山羊(♂),采集耳组织并分离培养成纤维细胞,以其为供体细胞,利用体细胞克隆技术开展超细型二狼山绒山羊的扩繁研究,通过克隆羊体重、羊绒细度、繁殖性能等指标评估该技术的可行性。【结果】试验成功建立了稳定的山羊卵母细胞体外成熟培养及体细胞克隆技术体系,卵母细胞成熟率为44.8%,孤雌激活卵裂率、囊胚率分别为83.0%、22.4%;二狼山绒山羊克隆胚的卵裂率、囊胚率分别为77.8%、13.7%。获得5只克隆羊个体,其生长发育正常,二狼山绒山羊母羊自然交配共获得16只健康的F1代个体,具有正常繁殖能力。克隆个体羊绒细度极显著低于同龄对照个体(P＜0.01)。【结论】本研究通过体细胞克隆技术成功克隆了超细型二狼山绒山羊个体,其具备正常生产性能和繁殖性能,并保留了超细羊绒的特性,为地方优良羊种质资源扩繁和新品种培育应用研究奠定了基础。

关键词: 二狼山绒山羊; 体细胞克隆; 羊绒细度; 繁殖性能

Abstract: 【Objective】 The cloned individual of superfine-type Erlangshan cashmere goat was produced by somatic cell cloning technology,which provided a theoretical basis for the application of this technology in the expansion of local high-quality sheep breed resources and the breeding of new varieties.【Method】 A 4-year old Erlangshan cashmere goat with a cashmere fineness of 13.89 μm was selected.The ear tissue was collected and cultured as fibroblasts,which were used as the donor cells.Somatic cell cloning technology was used to expand the propagation of superfine-type Erlangshan cashmere goat.The feasibility of the technique was evaluated by weight,fineness of cashmere and reproductive performance of cloned sheep.【Result】 The stable system of goat oocyte maturation culture in vitro and somatic cell cloning was successfully established.The oocyte maturation rate was 44.8%,and the cleavage rate and blastocyst rate of parthenogenesis was 83.0% and 22.4%,respectively.The cleavage rate and blastocyst rate of the cloned embryos of Erlangshan cashmere goat was 77.8% and 13.7%,respectively.5 cloned sheep were obtained with normal growth and development,and 16 healthy F1 generation individuals were obtained by natural mating with Erlangshan ewes,with normal reproductive ability.The cashmere fineness of cloned individuals was significantly lower than that of control individuals of the same age (P＜0.01).【Conclusion】 Superfine-type Erlangshan cashmere goat individuals was successfully cloned by somatic cell cloning technology,which had normal production and reproductive performance,and retained the characteristics of superfine cashmere,laid a foundation for the expansion of native excellent sheep germplasm resources and the application of new varieties breeding.

Key words: Erlangshan cashmere goat; somatic cell cloning; cashmere fineness; reproductive performance

中图分类号:

S813.3

卿玉波, 程雯, 角德灵, 刘城, 郝志强, 郭建雄, 李钰莹, 张筱琳, 李臻, 李效宇, 王文, 魏红江. 体细胞克隆技术扩繁超细型二狼山绒山羊的应用研究[J]. 中国畜牧兽医, 2024, 51(4): 1420-1427.

QING Yubo, CHENG Wen, JIAO Deling, LIU Cheng, HAO Zhiqiang, GUO Jianxiong, LI Yuying, ZHANG Xiaolin, LI Zhen, LI Xiaoyu, WANG Wen, WEI Hongjiang. Study on the Application of Propagation of Superfine-type Erlangshan Cashmere Goat by Somatic Cell Cloning Technology[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1420-1427.

参考文献

[1] CAI Y,FU W,CAI D,et al.Ancient genomes reveal the evolutionary history and origin of cashmere-producing goats in China[J].Molecular Biology and Evolution,2020,37(7):2099-2109.
[2] YANG F,LIU Z,ZHAO M,et al.Skin transcriptome reveals the periodic changes in genes underlying cashmere (ground hair) follicle transition in cashmere goats[J].BMC Genomics,2020,21(1):392.
[3] ZHANG Y,ZHANG D,XU Y,et al.Selection of cashmere fineness functional genes by translatomics[J].Frontiers in Genetics,2022,12:775499.
[4] JIN M,CAO Q,WANG R,et al.Molecular characterization and expression pattern of a novel keratin-associated protein 11.1 gene in the Liaoning cashmere goat (Capra hircus)[J].Asian-Australasian Journal of Animal Sciences,2017,30(3):328-337.
[5] WUDUBALA,LIU B,GAOWA MA Y J,et al.Comparative analysis of the main production performance between Chinese and Mongolian cashmere goat[J].Animal Husbandry and Feed Science,2016,37:19-22.
[6] YIN T,WANG J,XIANG H,et al.Dynamic characteristics of the mitochondrial genome in SCNT pigs[J].Biological Chemistry,2019,400(5):613-623.
[7] VAZQUEZ-AVENDANO J R,CORTEZ-ROMERO C,BRAVO-VINAJA A,et al.Reproduction of sheep through nuclear transfer of somatic cells:A bibliometric approach[J].Animals (Basel),2023,13(11):1839.
[8] WU H,ZHOU W,LIU H,et al.Whole-genome methylation analysis reveals epigenetic variation between wild-type and nontransgenic cloned,ASMT transgenic cloned dairy goats generated by the somatic cell nuclear transfer[J].Journal of Animal Science and Biotechnology,2022,13(1):145.
[9] JIN M,ZHAO L,YANG H,et al.A long non-coding RNA essential for early embryonic development improves somatic cell nuclear transfer somatic cell nuclear transfer efficiency in goats[J].Reproduction,2023,166(4):285-297.
[10] LIU F,LIU J,YUAN Z,et al.Generation of GTKO Diannan Miniature pig expressing human complementary regulator proteins hCD55 and hCD59via T2A peptide-based bicistronic vectors and SCNT[J].Molecular Biotechnology,2018,60(8):550-562.
[11] SHEN Y,XU K,YUAN Z,et al.Efficient generation of P53 biallelic knockout Diannan Miniature pigs via TALENs and somatic cell nuclear transfer[J].Journal of Translational Medicine,2017,15(1):224.
[12] ZHAO H,LI Y,WIRIYAHDAMRONG T,et al.Improved production of GTKO/hCD55/hCD59 triple-gene-modified Diannan Miniature pigs for xenotransplantation by recloning[J].Transgenic Rssearch,2020,29(3):369-379.
[13] WANG J,KHAN S U,CAO P,et al.Construction of PIK3C3 transgenic pig and its pathogenesis of liver damage[J].Life (Basel),2022,12(5):630.
[14] LI H,CHENG W,CHEN B,et al.Efficient generation of P53 biallelic mutations in Diannan Miniature pigs using RNA-guided base editing[J].Life (Basel),2021,11(12):1417.
[15] YU H,LONG W,ZHANG X,et al.Generation of GHR-modified pigs as Laron syndrome models via a dual-sgRNAs/Cas9 system and somatic cell nuclear transfer[J].Journal of Translational Medicine,2018,16(1):41.
[16] ZHU X X,ZHAN Q M,WEI Y Y,et al.CRISPR/Cas9-mediated MSTN disruption accelerates the growth of Chinese Bama pigs[J].Reproduction in Domestic Animals,2020,55(10):1314-1327.
[17] YAO J,ZENG H,ZHANG M,et al.OSBPL2-disrupted pigs recapitulate dual features of human hearing loss and hypercholesterolaemia[J].Journal of Genetics and Genomics,2019,46(8):379-387.
[18] ZHU X X,ZHONG Y Z,GE Y W,et al.Generation of transgenic-cloned Huanjiang Xiang pigs systemically expressing enhanced green fluorescent protein[J].Reproduction in Domestic Animals,2018,53(6):1546-1554.
[19] LI H H,WANG G,HAO Z Q,et al.Generation of biallelic knock-out sheep via gene-editing and somatic cell nuclear transfer[J].Scientific Reports,2016,6:33675.
[20] LEIBO S P,SZTEIN J M.Cryopreservation of mammalian embryos:Derivation of a method[J].Cryobiology,2019,86:1-9.
[21] KHAN S,TALI M,KHAN A,et al.Comparison of efficiency of in vitro cloned sheep embryo production by conventional somatic cell nuclear transfer and handmade cloning technique[J].Reproduction in Domestic Animals,2018,53(2):512-518.
[22] YANG X,SMITH S L,TIAN X C,et al.Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning[J].Nature Genetics,2007,39(3):295-302.
[23] SRIRATTANA K,KANEDA M,PARNPAI R.Strategies to improve the efficiency of somatic cell nuclear transfer[J].International Journal of Molecular Sciences,2022,23(4):1969.
[24] YU T,MENG R,SONG W,et al.ZFP57 regulates DNA methylation of imprinted genes to facilitate embryonic development of somatic cell nuclear transfer embryos in Holstein cows[J].Journal of Dairy Science,2023,106(1):769-782.
[25] WANG Y,LIU Q,KANG J,et al.Overexpression of PGC7 in donor cells maintains the DNA methylation status of imprinted genes in goat embryos derived from somatic cell nuclear transfer technology[J].Theriogenology,2020,151:86-94.
[26] 中国纤维质量监测中心.羊毛纤维直径试验方法——投影显微镜法:GB/T 10685—2007[S].2007. CHINA FIBER QUALITY MONITORING CENTER.Test method for diameter of wool fibers—Projection microscope method:GB/T 10685—2007[S].2007.(in Chinese)
[27] 王德军,陈炜,于洪川.山羊绒细度的不同仪器测定方法的对比试验研究[J].宁夏农学院学报,2002,2:27-29. WANG D J,CHEN W,YU H C.Comparative experimental study on different instrument measurement methods for cashmere fineness[J].Journal of Ningxia Agricultural College,2002,2:27-29.(in Chinese)

体细胞克隆技术扩繁超细型二狼山绒山羊的应用研究

Study on the Application of Propagation of Superfine-type Erlangshan Cashmere Goat by Somatic Cell Cloning Technology

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