一步3’RACE快速构建鸡MnSOD全长cDNA克隆Rapid |
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引用本文: | 卜友泉,罗绪刚,刘彬,李素芬.一步3’RACE快速构建鸡MnSOD全长cDNA克隆Rapid[J].遗传,2004,26(4):519-521. |
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作者姓名: | 卜友泉 罗绪刚 刘彬 李素芬 |
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作者单位: | 中国农业科学院畜牧研究所,北京 100094 Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100094 China |
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基金项目: | 国家杰出青年科学基金(39925028),北京市自然科学基金(6992022),国家人事部留学回国基金(7)资助~~ |
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摘 要: | 本研究尝试将触减 PCR与3’ cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术进行结合,仅用一条特异性引物和一条通用引物,成功地实现了从3’末端cDNA库对鸡含锰超氧化物歧化酶(manganese-containing superoxide dismutase,MnSOD)全长cDNA的一步3’RACE快速构建。与常规使用的末端PCR或亚克隆方法相比,该法具有快速、省时、经济和特异性好的优点。Abstract: RACE(rapid amplification of cDNA ends) is a popular technique to rapidly obtain the full-length cDNA. After obtaining the 3’cDNA and 5’cDNA fragments with a overlapped region by 3’RACE and 5’RACE, the full-length cDNA could be generated by end-to-end PCR or subcloning. In this study, 3’RACE combined with touch-down PCR was successfully used for the rapid construction of full-length MnSOD cDNA of chickens. Compared with the conventional end-to-end PCR or subcloning, this method, called one-step 3’RACE, is fast, economical and highly specific. It especially fits the rapid construction of full-length cDNA by RACE method.
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关 键 词: | RACE 触减PCR MnSOD RACE cDNA Key words |
文章编号: | 0253-9772(2004)04-0519-03 |
修稿时间: | 2003年5月26日 |
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