| Accession number;04A0070916 |
| Title;Cloning of the rpoZ gene encoding RNA polymerase .OMEGA. subunit from a deep-sea piezophilic bacterium |
| Author;NAKASONE K(Kinki Univ.) |
Journal Title;Research Reports of the Faculty of Engineering, Kinki University
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Journal Code:G0851A
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ISSN:0386-491X
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VOL.;NO.37;PAGE.33-41(2003)
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| Figure&Table&Reference;FIG.4, REF.11 |
| Pub. Country;Japan |
| Language;English |
| Abstract;We have cloned the rpoZ gene, encoding RNA polymerase .OMEGA. protein, by PCR approach from the deep-sea piezophilic and psychrophilic bacterium, Shewanella violacea strain DSS12. The cloned gene, 285bp in length, was found to encode a protein consisting of 93 amino acid residues with a molecular mass of 10,227Da. 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 Haemophilus influenzae (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 Ni2+ affinity column. The result of SDS-PAGE during purification suggests that a chimeric RNA polymerase (.ALPHA.2.BETA..BETA.'.SIGMA.70.OMEGA.) was assembled in E. coli. (author abst.) |
