组织特异过表达FST基因促进阿尔巴斯绒山羊肌肉生长  

Tissue-specific Overexpression of FST Promote Muscle Growth in Albas Cashmere Goats

作　　者：张驹 王玮 王珂 杨楠 马建飞 路大同 仓明 ZHANG Ju;WANG Wei;WANG Ke;YANG Nan;MA Jian-fei;LU Da-tong;CANG Ming(State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock,Inner Mongolia University,Hohhot 010070,China)

机构地区：[1]内蒙古大学省部共建草原家畜生殖调控与繁育国家重点实验室

出　　处：《内蒙古大学学报（自然科学版）》2019年第4期353-363,共11页Journal of Inner Mongolia University：Natural Science Edition

摘　　要：高产优质肉用家畜新品种的培育是家畜育种的工作目标之一,体细胞核移植技术和基因编辑技术的发展为新品种家畜的培育提供了快速便捷的新方式.本研究将骨骼肌特异表达卵泡抑制素(Follistatin,FST)的pCDsRed2-SP-FST重组质粒转染山羊胎儿成纤维细胞,通过G418筛选获得稳定转染的阳性供体细胞系,将供体细胞注入去核成熟卵母细胞获得FST基因编辑重构胚胎425个,胚胎移植到59个受体母羊,出生羔羊4只,其中1只羔羊存活至今.利用PCR、Southern Blot、Western Blot和qPCR等技术在分子生物学水平鉴定该羊羔,发现FST基因被成功插入到山羊基因组中并可在肌肉组织中过表达,而且FST基因的过表达可使肌肉标志基因MSTN和BMP4的表达增强.羔羊健康分析表明转FST基因绒山羊各项生理指标与野生型相比并无明显差异,但FST基因的过表达增加了肌纤维的直径和密度.该FST基因过表达绒山羊的成功获得为高产优质肉用家畜新品种的培育奠定了研究基础.Breeding of livestock with fast growth and high-quality meat traits is a major purpose of animal breeding.The development of somatic cell nuclear transfer(SCNT)technology and gene editing technology provides a fast and convenient route for breeding.In this study,pCDsRed2-SPFST recombinant plasmid with skeletal muscle specifically expressing follistatin(FST)feature was transfected into goat fetal fibroblasts.Stable transfected positive donor cell lines were obtained by G418 screening.Then 425 SCNT embryos were constructed and transferred to 59 recipient ewes.Four recipients were pregnant and delivered four lambs,in which one survived.The survived one was evaluated by PCR,Southern Blot,Western Blot and qPCR.The results showed FST gene was successfully inserted into the genome and over-expressed in lamb muscle tissue.Moreover,the increased expression of MSTN and BMP4 was induced by FST over-expression.The physiological evaluation showed that there were no significant difference between FST goat and control.However,the overexpression of FST gene increased the diameter and density of muscle fibers in the lamb.The successful obtaining of FST over-expressed cashmere goats provided valuable basic for breeding new livestock breeds with high meat yield and quality.

关 键 词：体细胞核移植 卵泡抑制素 骨骼肌特异表达载体 肌肉 转FST基因绒山羊 

分 类 号：S814.8[农业科学—畜牧学] Q786[农业科学—畜牧兽医]