体细胞核移植技术生产转基因猪胚胎的研究  

A Study of Transgenic Pig Embryos Produced by Somatic Cell Nuclear Transfer

作　　者：华再东[1] 刘西梅[1] 毕延震[1] 华升 郑新民[1] 

机构地区：[1]湖北省农业科学院,动物胚胎工程及分子育种湖北省重点实验室,湖北武汉430064 [2]武汉金鹰牧业有限公司,湖北武汉430209

出　　处：《华南农业大学学报》2013年第3期394-398,421,共6页Journal of South China Agricultural University

基　　金：国家转基因生物新品种培育重大专项(2013ZX08006-002;2013ZX08010-3;2013ZX08006-003);湖北省农业科技创新中心课题(2011-620-001-003);湖北省自然科学基金(2010CDA120);湖北省国际科技合作项目(2009BFA012)

摘　　要：以转染绿色荧光蛋白基因的猪胎儿成纤维阳性细胞作为体细胞核移植的核供体,体外成熟卵母细胞为核移植受体构建绿色荧光蛋白转基因克隆猪胚胎,研究供核细胞的处理、注核部位及重构胚融合/激活时间对转GFP克隆胚早期发育的影响.结果显示,胎儿成纤维细胞血清饥饿与非饥饿培养10 d处理组,采用卵周间隙核移植重构胚的卵裂率(82.35%和79.07%)差异不显著(P>0.05);体外培养42～44 h卵母细胞进行胞质内和卵周间隙注核的重构胚胎,其卵裂率(81.11%和76.80%)差异不显著(P>0.05);将卵周间隙注射法构建重构胚在0～1 h,2～4 h和6～8 h进行融合/激活操作,前2组重构胚卵裂率(75.61%和83.07%)无显著差异(P>0.05),但显著高于第3组(60.00%)的卵裂率(P<0.05).In the present study,green fluorescent protein（GFP） transgenic embryos were reconstructed by nuclear transfer of GFP positive cells into enucleated in vitro matured oocytes.The development of reconstructed embryos both in vitro was observed,and GFP expression was checked as well.This research was conducted to study the processing of nuclear cells,the method of nuclear transfer and fusion / activation on early development in vitro of reconstructed embryos.The results showed that there were no significant differences in the cleavage rates between serum hunger and not hunger training 10 d using injection ways of perirenal space of eggs,and the cleavage rate of reconstructed embryos with intracytoplasmic injection（81.11%） was not significantly higher than that of the perirenal space of eggs（76.80%）.After activation treatment,the rate of cleavage were assessed on 2 days,respectively.Although there were no significant differences in the cleavage rates of the first two treatment groups（75.61% and 83.07%）,the developmental rate was significantly higher than that of the third group（60.00%）.

关 键 词：供核细胞 融合激活时间 转基因重构胚 猪 

分 类 号：S864.3[农业科学—野生动物驯养]