深海微生物由来RNAポリメラーゼωサブユニットをコードするrpoZ遺伝子のクローニング

概要

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We have cloned the rpoZ gene, encoding RNA polymerase ω protein, by PCR approach from the deep-sea piezophilic and psychrophilic bacterium, Shewanella violacea strain DSS12. The cloned gene, 285 bp in length, was found to encode a protein consisting of 93 amino acid residues with a molecular mass of 10,227 Da. Significant homology was evident comparing the rpoZ protein of S. violacea with that of Escherichia coli K-12 (71% identity), Vibrio cholerae (70% identity) and Haemophilia influenzas (61% identity). Phylogenetic analysis of RpoZ proteins of several bacteria suggested that S. violacea is independent from other bacteria. We constructed expression plasmid to overproduce the RpoZ protein and transformed into E. coli JM109 as a host of overproduction. Upon induction, the recombinant protein encoded by plasmid pQrpoZ was overexpressed and purified using Ni^<2+> affinity column. The result of SDS-PAGE during purification suggests that a chimeric RNA polymerase (α_2ββ'σ^<70>ω) was assembled in E. coli
近畿大学の論文
2003-12-20