摘要
介绍了一个简便的DNA改组操作程序 .首先利用PCR扩增了两段具有高度序列同源性的 170 0bp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Mg2 +存在的条件下 ,用DNaseⅠ切割成 10～5 0bp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 .
A facile DNA shuffling protocol was introduced. Two genes of about 1 700 bp were obtained by PCR from two different individual templates, separately, which shared over 93% homology. After mixed with equimolar of each, these two genes were further cut randomly by DNaseⅠ into small fragments of 10～50 bp under the existence of Mg 2+ . These small fragments were successfully re assembled 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.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2000年第6期655-657,共3页
Progress In Biochemistry and Biophysics
基金
(1986~1995)国家高技术发展计划项目
(1996~1998)国家自然科学基金资助项目
