安徽农业科学Issue(19)：10166-10169,4.
水牛受精卵胞质内注射转基因的初步研究
Study on Transgenesis by Cytoplasmic Injection of Exogenous DNA into Buffalo Zygote
摘要
Abstract
[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of buffalo. [Method ] Buffalo oocytes were randomly divided into two groups 20 -22 h after in vitro maturation. One group of oocytes was introduced with about 7. 5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7 - 10 h or 18 - 20 h after in vitro fertilization (IVF); the other group of ooctyes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear ECFP fragment by cytoplasmic injection ( generally called ICSI-Mediated Gene Transfer, ICSI-Tr). Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blas-tocyst gene expression efficiency in IVF group showed no significant difference compared with that in ICSI-Tr group (P >0.05). In addition, the cleavage rate and early embryonic gene expression efficiency in IVF group were significantly higher with injection at 7 - 10 h pos1 IVF than 18 - 20 h post IVF (P < 0.05 ). [Conclusion ] These results indicate that transgenie buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7 - 10 h post IVF.关键词
水牛/受精卵/胞质内注射/转基因Key words
Buffalo/ Zygote/ Cytoplasmic injection/ Transgenesis分类
农业科技引用本文复制引用
陈自洪,崔奎青,孟凡丽,刘玉兵,王丹,陆凤花,石德顺..水牛受精卵胞质内注射转基因的初步研究[J].安徽农业科学,2012,(19):10166-10169,4.基金项目
国家高技术研究发展计划(863)项目(2007AA100505) （863）
国家转基因重大专项(2008ZX08007-003). （2008ZX08007-003）
