用于创建单交换文库的体外模板改变PCR：以苏云金芽孢杆菌Cry2A毒素为例的研究
In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins.
作者信息
Shu Changlong, Zhou Jianqiao, Crickmore Neil, Li Xianchun, Song Fuping, Liang Gemei, He Kanglai, Huang Dafang, Zhang Jie
机构信息
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, P. R. China.
School of Life Sciences, University of Sussex, Falmer, Brighton, UK.
出版信息
Sci Rep. 2016 Apr 21;6:23536. doi: 10.1038/srep23536.
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.
在进化过程中，重复的旁系同源基因之间产生单交换嵌合体是增加多样性的一个已知过程。比较具有一个或两个交换的同源重组嵌合体的特性也是分析序列变异与功能之间关系的有效策略。然而，尚未建立完善的体外方法来创建单交换文库。在此，我们展示了一种体外模板改变聚合酶改变反应，该反应已被开发用于产生此类文库。我们将该方法应用于来自苏云金芽孢杆菌的两个密切相关的毒素基因，并创建了具有不同特性的嵌合体，这有助于我们理解这些毒素如何区分不同昆虫物种。
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