一种新的等温单向生长基因合成法初步探索  

Isothermal unidirectional elongation method of gene synthesis

作　　者：林继伟[1] 张晓东[1] 曹雪雁[1] 胡钧[1,2] 

机构地区：[1]上海交通大学生命科学技术学院,Bio-X生命科学研究中心 [2]中国科学院上海应用物理研究所,上海201800

出　　处：《遗传》2007年第6期765-770,共6页Hereditas（Beijing)

基　　金：国家自然科学基金(编号:10335070;10674147);上海市科委创新团队项目和纳米专项(编号:03DZ14025;0652NM006)资助~~

摘　　要：基因合成正日益成为基因获取的一种有效手段。两种常用的基因合成方法是基于PCR的基因合成(P法)和基于连接酶的基因合成(L法)。这两种方法都不是独立于所合成基因的序列内容的。文章首次提出了一种新的基因合成策略-等温单向生长法(Isothermal Unidirectional Elongation Method,IUEM),在3种酶的作用下,DNA链在等温状态下单向串行延伸,直到获得目标基因。由于引物设计成特殊的发夹结构,该方法可以做到合成过程独立于或部分独立于目标基因的序列内容。本文探索了该策略的实验可行性,检测了影响基因合成的各种因素,并利用该方法成功地合成了一段254bp和一段300bp的DNA序列。Gene synthesis is gaining popularity as a versatile means of cloning genes. The prevailing synthesis methods are PCR-based and llgase-based gene synthesis. Both of them are not independent of the sequence content of the target gene. In the present study, a new gene synthesis method-isothermal unidirectional elongation method （IUEM） is proposed, which involves the cooperation of three enzymes, and is independent or partially independent of the target gene sequence by the specially designed oligonucleotides that form hairpins. The synthesis process is isothermal and serial in IUEM. Several factors that affect the elongation were studied, and two DNA segments of length 254 bp and 300 bp were synthesized with IUEM. The synthesized segments could work as building blocks for the synthesis of longer DNA sequences.

关 键 词：基因合成 等温 单向 发夹结构 

分 类 号：Q78[生物学—分子生物学]