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

Shu, Changlong, Zhou, Jianqiao, Crickmore, Neil, Li, Xianchun, Song, Fuping, Liang, Gemei, He, Kanglai, Huang, Dafang and Zhang, Jie (2016) In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins. Scientific Reports, 6. p. 23536. ISSN 2045-2322

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Abstract

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.

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
Subjects: Q Science > QR Microbiology
Q Science > QR Microbiology > QR0075 Bacteria
Depositing User: Neil Crickmore
Date Deposited: 27 Apr 2016 09:06
Last Modified: 08 Mar 2017 01:05
URI: http://sro.sussex.ac.uk/id/eprint/60628

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