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| 水牛受精卵胞质内注射转基因的初步研究 | |
| 引用本文： | 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. |
| 作者姓名： | 陈自洪 崔奎青 孟凡丽 刘玉兵 王丹 陆凤花 石德顺 |
| 作者单位： | 1. 广西大学亚热带生物资源保护利用国家重点实验室,广西南宁530004;广西壮族自治区人口和计划生育研究中心,广西南宁530021 2. 广西大学亚热带生物资源保护利用国家重点实验室,广西南宁,530004 3. 广西大学亚热带生物资源保护利用国家重点实验室,广西南宁530004;山东潍坊市人民医院生殖医学中心,山东潍坊261100 |
| 基金项目： | 国家高技术研究发展计划(863)项目，国家转基因重大专项 |
| 摘 要： | 目的]旨在探讨通过向水牛受精卵胞质内注射外源DNA实现转基因的可行性。方法]水牛卵母细胞体外成熟20～22 h后随机分为2组:①在体外受精7～10 h或18～20 h后向卵胞质内注入约7.5 pl 50μg/ml含线性EGFP片段的DNA溶液;②分别注入单个精子与约7.5 pl 50μg/ml含线性EGFP片段的DNA混合物,观察外源基因在胚胎发育过程中的表达情况。结果]受精卵胞质内注射的早期胚胎基因表达率、囊胚基因表达率与ICSI-Tr差异不显著(P0.05),且IVF 7～10 h时注射的分裂率、早期胚胎基因表达率均显著高于18～20 h(P0.05)。结论]水牛IVF受精卵胞质内注射外源基因能获得转基因胚胎,且IVF后7～10 h注射的效果优于IVF后18～20 h注射。 |
| 关 键 词： | 水牛 受精卵 胞质内注射 转基因 |
Study on Transgenesis by Cytoplasmic Injection of Exogenous DNA into Buffalo Zygote |
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| Institution: | CHEN Zi-hong et al(State Key Laboratory for Conservation and Utilization of Agricultural Bioresources in the Subtropics,Guangxi University,Nanning,Guangxi 530004) |
| 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 EGFP 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 blastocyst 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 post IVF than 18-20 h post IVF(P<0.05).Conclusion] These results indicate that transgenic 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. |
| Keywords: | Buffalo Zygote Cytoplasmic injection Transgenesis |
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