| 以“末端修饰法”构建人基因组的酵母人工染色体克隆 |
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| 引用本文： | 张小晓,刘国仰.以“末端修饰法”构建人基因组的酵母人工染色体克隆[J].中国医学科学院学报,1997,19(3):163-170. |
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| 作者姓名： | 张小晓 刘国仰 |
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| 作者单位： | 复旦大学遗传工程国家重点实验室遗传研究所!上海(张小晓，邓余，杨玉美，罗会元)，200433(余龙，刘国仰，黄尚志)，中国医学科学院中国协和医科大学基础医学研究所(杨焕明)，20043(赵寿元) |
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| 基金项目： | 国家863高科技基金!863-102-19-5，国家自然科学基金!39180005 |
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| 摘 要： | 采用“末端修饰法”修饰酵母人工染色体（YAC）载体与基因组插入片段的酶切末端，并优化各个实验参数，成功地得到了4000多个YAC片段的克隆群，转化率为400～500转化子／μgDNA。经随机抽样初步鉴定：80％的阳性克隆能检出人工染色体，插入片段的大小范围为150～750kb，平均为450kb。载体连接实验检测表明，末端修饰法能有效防止“共连接”的发生。
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| 关 键 词： | 酵母人工染色体克隆 酵母人工染色体嵌合体 末端修饰 |
Construction of Human Genomic YAC Clones with "Terminal Modification |
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| Zhang Xiaoxiao, Yu Long Deng, Yu Liu, Guoyang Yang, Yumei,et al.Construction of Human Genomic YAC Clones with "Terminal Modification[J].Acta Academiae Medicinae Sinicae,1997,19(3):163-170. |
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| Authors: | Zhang Xiaoxiao Yu Long Deng Yu Liu Guoyang Yang Yumei |
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| Institution: | State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, Shanghai. |
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| Abstract: | A human YAC genomic pool with more than 4 000 clones was constructed by means of "terminal modification" of YAC vector and genomic inserts, as well as optimization of rele-vant experimental pararneters. The transformation rate is approximately 4OO~5OO transfor-mants per pg DNA. Preliminary characterization of randomly sampled clones demonstrates that YAC-specific inserts, in the average size of 450 kb and ranging 150~ 750 kb,were de-tected from 8O% of the clones. The vector ligation experiment indicates the efficient reduc-tion of "coligation" by tertninal modification. |
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| Keywords: | YAC cloning chimerism terminal modification |
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