Bacterial ArtA protein specifically binds to the internal region of IS1 in vitro英文文献资料.docVIP

Advances in Bioscience and Biotechnology, 2012, 3, 869-875 ABB /10.4236/abb.2012.37108 Published Online November 2012 (http://www.SciRP.org/journal/abb/) Bacterial ArtA protein specifically binds to the internal region of IS1 in vitro Sachiko Matsutani Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan Email: sachiko@nihs.go.jp Received 12 August 2012; revised 21 September 2012; accepted 24 October 2012 ABSTRACT lates transcription [5]. The genetic results does not show whether the ArtA protein binds directly to the IS1 inter- nal region. The amino acid sequence of ArtA, which consists of 104 residues, has no detectable match with the known protein domain families in Pfam database (unpublished data). It is significant to examine the DNA- binding ability of ArtA in vitro. So far, I had attempted to clone artA to various expression vectors and produce ArtA in various host strains. However, the production was never observed on SDS-polyacrylamide gels. When Wu and Ippen-Ihler [7] found the artA gene in the tra region of the Escherichia coli F factor as a gene of un- known function, its expression was examined: The prod- uct seems not to be expressed in E. coli maxicells, but the expression potential of the promoter and transla- tional start site for artA are in upstream region. artA is AT rich and has many rare codons of E. coli (unpub- lished data), and thus the gene product might not be de- tected. Recently, many genes have artificially synthe- sized with codons optimized for the expression host, which led to higher protein production [8]. The internal region of bacterial translocatable IS1 acts as a cis-element to stimulate transcription from the various promoters located upstream. The product of the artA gene is genetically shown to stimulate transcription with the cis-element. Here, a codon- optimized artA