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一种改进的基因合成方法
引用本文:王文凯,蓝东明,王永华,杨博. 一种改进的基因合成方法[J]. 安徽农业科学, 2011, 39(18): 10735-10737
作者姓名:王文凯  蓝东明  王永华  杨博
作者单位:华南理工大学生物科学与工程学院,广东,广州,510006;华南理工大学轻工与食品学院,广东,广州,510640
基金项目:“酶工程关键技术与产业化”(2009A1-E021)
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(ImprovedoverlapextensionPCRmethod)法。[结果]拆分的单链寡核苷酸长度为60nt左右,重叠区碱基数13~17bp,Tm值为40-50℃。IOEP法具体步骤如下:把拆分的寡核苷酸组装成500bp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%o,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。

关 键 词:基因合成  IOEP  化学合成成本率

An Improved Method for Gene Synthesis
WANG Wen-kai et al. An Improved Method for Gene Synthesis[J]. Journal of Anhui Agricultural Sciences, 2011, 39(18): 10735-10737
Authors:WANG Wen-kai et al
Affiliation:WANG Wen-kai et al(School of Bioscience and Bioengineering,South China University of Technology,Guangzhou,Guangdong 510006)
Abstract:[Objective]The research aimed to set up a simple,cheap and accurate gene synthesis method.[Method]The existing overlap-extension synthesis method was improved.And an improved overlap extension PCR(IOEP)methodwas established.[Result]The splited oligonucleotides length was about 60 nt.The base number in overlapped region was 13-17 bp,Tm was 40-50 ℃.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.
Keywords:Gene synthesis  IOEP  Cost ratio of chemical synthesis  
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