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学科主题: 生命有机化学
题名: 一个简便的DNA改组(DNA shuffling)操作程序
其他题名: A facile DNA shuffling protocol
作者: 周佳海 ; 陈海宝
通讯作者: 陈海宝
刊名: 生物化学与生物物理进展
发表日期: 2000-01-01
卷: 27, 期:6, 页:655-657
收录类别: SCI
部门归属: 中国科学院上海有机化学研究所生命有机国家开放实验室
摘要: 介绍了一个简便的DNA改组操作程序.首先利用PCR增了两段具有高度序列同源性的1伽bb左右的基因片段,两者相比较同源性大于93%.然后将其等量混合后,在Mg~2存在的条件下,用DNaseI切割成10一50 bp的小片段.这些小片段在不外加引物的前提下,利用PcR反应进行重聚,再将重聚物经过两轮正常的PCR扩增,获得了与原来片段大小相当的基因片段.这一技术有利于从一组序列同源性程度较高的基因库构建随即嵌合基因·
英文摘要: A facile DNA shuffling protocol was introduced. Two genes of about 1 700 bp were obtained by PCR fromtwo different individual templates, separately, which shared over 93% homology. After mixed with equimolar of each, these two genes were further cut randomly by Dnase I into small fragments of 10~50 bp under the existence of Mg~2+. These small fragments were successfully reassembled to form full genes with original size by one round of PCR without any external primers and two other rounds of normal PCR amplification. This shuffling protocol may help to construct chimera genes from a family of genes with high sequence homology. R
语种: 中文
内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/25743
Appears in Collections:生命有机化学国家重点实验室_期刊论文

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Recommended Citation:
周佳海,陈海宝. 一个简便的DNA改组(DNA shuffling)操作程序[J]. 生物化学与生物物理进展,2000,27(6):655-657.
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