In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins

文献类型： 外文期刊

第一作者： Zhou, Jianqiao

作者： Zhou, Jianqiao;Li, Xianchun;Song, Fuping;Liang, Gemei;He, Kanglai;Zhang, Jie;Crickmore, Neil;Huang, Dafang

作者机构：

期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）

ISSN： 2045-2322

年卷期： 2016 年 6 卷

页码：

收录情况： SCI

摘要： During evolution the creation of single crossover chimeras between duplicated paralogous genes is a known process for increasing diversity. Comparing the properties of homologously recombined chimeras with one or two crossovers is also an efficient strategy for analyzing relationships between sequence variation and function. However, no well-developed in vitro method has been established to create single-crossover libraries. Here we present an in vitro template-change polymerase change reaction that has been developed to enable the production of such libraries. We applied the method to two closely related toxin genes from B. thuringiensis and created chimeras with differing properties that can help us understand how these toxins are able to differentiate between insect species.

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