Purification and Properties of the Plasmid Maintenance Proteins from the Borrelia burgdorferi Linear Plasmid lp17
| Autor: | George Chaconas, Jan Deneke |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Genetics
Plasmid partitioning Molecular Sequence Data DNA replication Genetics and Molecular Biology Gene Expression Regulation Bacterial Biology medicine.disease_cause biology.organism_classification Microbiology Plasmid maintenance law.invention Plasmid Bacterial Proteins law Borrelia burgdorferi medicine Recombinant DNA Amino Acid Sequence Molecular Biology Escherichia coli Peptide sequence Plasmids |
| Zdroj: | Journal of Bacteriology. 190:3992-4000 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.00057-08 |
| Popis: | The Lyme disease spirochete Borrelia burgdorferi carries more plasmids than any other bacterium, many of which are linear with covalently closed hairpin ends. These plasmids have also been referred to as mini-chromosomes and essential genetic elements and are integral components of its segmented genome. We have investigated two plasmid maintenance proteins, BBD14 (the replication initiator) and BBD21 (a presumptive ParA orthologue), encoded by the linear plasmid lp17; these proteins are representatives of paralogous families 62 and 32, respectively. We have purified recombinant 6-his-BBD21 and shown it possesses an ATPase activity. 6-his-BBD14 initially could not be overexpressed in Escherichia coli by itself. It was only effectively overproduced in recombinant form through coexpression with other B. burgdorferi proteins and codon optimization. Although the mechanism for increased production through coexpression is not clear, this method holds promise for expression and purification of other B. burgdorferi proteins, a number of which have remained recalcitrant to purification from E. coli . Finally, we present evidence for the physical interaction of BBD14 and BBD21, a feature suggesting that BBD21 and the paralogous family 32 proteins are more likely involved in DNA replication than functioning as simple ParA orthologues as previously surmised based upon sequence homology. Such a role would not preclude a function in plasmid partitioning through interaction with the replication initiator. |
| Databáze: | OpenAIRE |
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