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  • 标题:In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins
  • 本地全文:下载
  • 作者:Changlong Shu ; Jianqiao Zhou ; Neil Crickmore
  • 期刊名称:Scientific Reports
  • 电子版ISSN:2045-2322
  • 出版年度:2016
  • 卷号:6
  • 期号:1
  • DOI:10.1038/srep23536
  • 语种:English
  • 出版社:Springer Nature
  • 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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