The possibilities of using plasmid ORF clone library for ybeY gene compensation studies in Escherichia coli
Date
2020
Authors
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Journal ISSN
Volume Title
Publisher
Tartu Ülikool
Abstract
The YbeY, encoded by the highly conserved ybeY gene, is a multi-functional protein in Escherichia coli. The processing of the 3′ terminus of 16S rRNA is its most important function, which has a direct effect on the biogenesis of ribosomes. The deletion of ybeY leads to phenotypic and translational defects. YbeY has an extensive network of interactions with other proteins, many of which are still not fully understood. ORF library can be used as a powerful biological tool to detect interactions between proteins. Two different selection strategies were applied using two pooled Gateway® entry clone libraries. Fully pooled library containing all genes represented in the library, and partially pooled library with the lack of some library genes, were electroporated into ybeY null mutants. According to the specificity of each pooled library, the growth selections and PCR methods for genes detecting were different. Results have shown that cultures growth improves during selection. Several genes, whose amount prevails over others in the last selection cycles, were sequenced.
Description
Keywords
YbeY, ORF library, selection, Gateway® entry clone library