摘要
[目的]建立一种简便、廉价、准确的基因合成方法。[方法]对现有的重叠延伸合成方法进行了改进,提出一种IOEP(Improvedoverlap extension PCR method)法。[结果]拆分的单链寡核苷酸长度为60 nt左右,重叠区碱基数13～17 bp,Tm值为40～50℃。IOEP法具体步骤如下:把拆分的寡核苷酸组装成500 bp左右的中间片段,再将中间片段拼接成全长基因。利用IOEP法合成了10条基因,其平均化学合成成本率为1.291,平均错误率为1.031‰,优于其他合成方法。[结论]IOEP法是一种具有高准确率、低成本、重复性好的基因合成方法。
[ Objective ] The research aimed to set up a simple, cheap and accurate gene synthesis method. [ Method] The existing overlap -exten- sion synthesis method was improved. And an improved overlap extension PCR （IOEP）methodwas established. [ Result] The splited oligonucleoti- des length was about 60 nt. The base number in overlapped region was 13 - 17 bp,Tm was 40 -50 ~C. The concrete steps of IOEP method were as follows：assembling splited oligonucleotides into 500 bp length of intermediate blocks and splicing the intermediate blocks into the full-length gene. Ten genes were synthesized by using IOEP method. The average cost ratio of chemical synthesis was 1. 291 and the average error rate was 1. 031‰. IOEP method was better than other methods. [ Conclusion ] IOEP method is a kind of gene synthesis method with high accuracy, lower cost and good repeatability.
出处
《安徽农业科学》
CAS
北大核心
2011年第18期10735-10737,共3页
Journal of Anhui Agricultural Sciences
基金
"酶工程关键技术与产业化"(2009A1-E021)
