Genome-wide identification of CodY target genes in Bacillus thuringiensis by in vitro binding analysis

Abstract:

[Objective] This study is aimed to establish an in vitro pull down method combining Avi-tag technology to screen and indentify the CodY targets in the whole genomes of Bacillus thuringiensis strain YBT-1520. The method offers a potential biocatalyst forscreening target sequences of transcription factors in vitro. [Methods] CodY proteins were labeled with biotin using Avi-tag technology. B. thuringiensis strain YBT-1520 genomes were enzymatically digested to 100-400 bp DNA fragments that had adapters with known sequences on both ends. Then, the target genes of CodY protein were screened by streptavidin biotin resin. [Results] The 46 targets of CodY were mainly involved in amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, transport, regulation, DNA replication repair, and two component regulatory systems. [Conclusion] Compared with chromatin immune co-precipitation technology in vivo, this pull down method is simple, cheap, fast and specific to biotin streptavidin system.

Key words:CodY;target genes;Avi-tag technology;Bacillus thuringiensis;biotin;regulation