| 电转染法电压优化与广西巴马小型猪转基因克隆胚胎的生产 |
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| 引用本文： | 黄勇,;朱向星,;全守能,;卢晟盛.电转染法电压优化与广西巴马小型猪转基因克隆胚胎的生产[J].广西农业生物科学,2014(2):348-356. |
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| 作者姓名： | 黄勇 ;朱向星 ;全守能 ;卢晟盛 |
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| 作者单位： | [1]广西大学亚热带农业生物资源保护与利用国家重点实验室,南宁530004; [2]广西大学动物科学技术学院,南宁530004 |
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| 基金项目： | 国家自然科学基金资助项目（31260553）; 广西硕士研究生创新课题（YCSZ2013003）共同资助 |
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| 摘 要： | 本试验旨在优化电转染电压并构建转基因体细胞,为生产转基因广西巴马小型猪打下技术储备。以新生广西巴马小型猪肾脏成纤维细胞和pEGFP-N1为材料、以细胞存活率和转染率为电转染效率评价指标,筛选最佳电压,然后用优化电压进行电转染并经G418抗性筛选获得转基因体细胞系,通过体细胞核移植技术获得转基因克隆胚胎。本研究筛选出最佳电压为120 V,用优化电压进行电转染并成功构建表达GFP的转基因细胞,最后通过体细胞核移植成功获得表达GFP的转基因克隆胚胎。结果表明优化的电转染电压可以高效构建广西巴马小型猪转基因体细胞,并可通过体细胞核移植生产转基因克隆胚胎。
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| 关 键 词： | 电转染 转基因 广西巴马小型猪 克隆 |
Optimization of the Voltage of Electrotransfection and Production of Bama Minipig Transgenic Cloned Embryos |
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| Institution: | Huang Yong , Zhu Xiangxing , Quan Shouneng , Lu Shengsheng( 1 State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, 530004; 2 College of Animal Science and Technology, Guangxi University, Nanning, 530004) |
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| Abstract: | The objective of this study was to optimize the voltage of electrotransfection for constructing porcine transgenic cells, which would be used to produce porcine transgenic cloned embryos via somatic cell nuclear transfer（SCNT）. The fibroblasts derived from newborn Guangxi Bama minipig kidney and pEGFP-N1 were used to optimize the voltage of electrotransfection, and transfected cells were suffered to G418 resistance screening for establishing transgenic cell line, which was employed to produce porcine transgenic cloned embryos via SCNT. It was showed that the best voltage of electrotransfection is 120 V, which was employed to establish Bama mimipig transgenic somatic cells, and then these transgenic cells via SCNT were used to produce porcine transgenic cloned embryos, which expressed green fluorescence during developmental stages in vitro. In conclusion, optimized voltage of electrotransfection could be efficiently employed to construct Bama mimipig transgenic somatic cells, which could be used to produce porcine transgenic cloned embryos via SCNT. |
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| Keywords: | Electrotransfection Transgenesis Guangxi Bama minipig Cloning |
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