Isolation and characterization of theRhodococcus opacusthiostrepton-inducible genestipALandtipAS: application for recombinant protein expression inRhodococcus

• DOI: 10.1111/j.1574-6968.2004.tb09675.x
• Publication Date: 2006-01-09T08:33:03Z
• Authors: Li Dong; Nobutaka Nakashima; Noriko Tamura; Tomohiro Tamura
• Keywords: DNA, Bacterial; 570; 0303 health sciences; Transcription, Genetic; Molecular Sequence Data; Gene Expression Regulation, Bacterial; Sequence Analysis, DNA; Thiostrepton; Recombinant Proteins; Artificial Gene Fusion; Culture Media; 03 medical and health sciences; Bacterial Proteins; Genes, Bacterial; Sequence Analysis, Protein; Rhodococcus; Cloning, Molecular; Promoter Regions, Genetic; Sequence Deletion

We cloned the Rhodococcus opacus (strain DSM 44193) tipA gene, which encodes two translation products, TipAL and TipAS. The gene products are homologous to the Streptomyces spp. TipAL and TipAS proteins, respectively. The tipA promoter is highly active and TipAS protein is predominantly accumulated in R. opacus cells when the inducer of transcription, thiostrepton, was presented in culture medium. We found that thiostrepton is also induced the expression of an endogenous TipA-family protein in Rhodococcus erythropolis (strain JCM3201). The minimal tipA promoter region was defined (57 bp) and the conserved nucleotide sequence of the putative TipAL protein binding site (TipA-box) was identified in that region. The tipA gene is presumed to be transcribed into a leaderless mRNA. We applied the tipA promoter successfully for recombinant protein expression in R. erythropolis cells.