Abstract
Cytophaga johnsonae displays many features that make it an excellent model of bacterial gliding motility. Unfortunately, genetic analyses of C. johnsonae, or any related gliding bacteria, were not possible because of a complete lack of selectable markers, cloning vectors, transposons, and convenient methods of gene transfer. As a first step toward a molecular analysis of gliding motility of C. johnsonae, we developed these genetic techniques and tools. Common broad-host-range plasmids and transposons did not function in C. johnsonae. We identified one Bacteroides transposon, Tn4351, that could be introduced into C. johnsonae on plasmid R751 by conjugation from Escherichia coli. Tn4351 inserted in the C. johnsonae genome and conferred erythromycin resistance. Tn-4351 insertions resulted in auxotrophic mutations and motility mutations. We constructed novel plasmids and cosmids for genetic analyses of C. johnsonae. These cloning vectors are derived from a small cryptic plasmid (pCP1) that we identified in the fish pathogen Cytophaga psychrophila D12. These plasmids contain the ermF (erythromycin resistance) gene from Tn4351 and a variety of features that facilitate propagation and selection in E. coli and conjugative transfer from E. coli to C. johnsonae.
Full Text
The Full Text of this article is available as a PDF (344.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson K. L., Salyers A. A. Genetic evidence that outer membrane binding of starch is required for starch utilization by Bacteroides thetaiotaomicron. J Bacteriol. 1989 Jun;171(6):3199–3204. doi: 10.1128/jb.171.6.3199-3204.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bagdasarian M., Lurz R., Rückert B., Franklin F. C., Bagdasarian M. M., Frey J., Timmis K. N. Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene. 1981 Dec;16(1-3):237–247. doi: 10.1016/0378-1119(81)90080-9. [DOI] [PubMed] [Google Scholar]
- Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
- Brenner D. J., Hollis D. G., Fanning G. R., Weaver R. E. Capnocytophaga canimorsus sp. nov. (formerly CDC group DF-2), a cause of septicemia following dog bite, and C. cynodegmi sp. nov., a cause of localized wound infection following dog bite. J Clin Microbiol. 1989 Feb;27(2):231–235. doi: 10.1128/jcm.27.2.231-235.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buu-Hoi A. Y., Joundy S., Acar J. F. Endocarditis caused by Capnocytophaga ochracea. J Clin Microbiol. 1988 May;26(5):1061–1062. doi: 10.1128/jcm.26.5.1061-1062.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheng Q., Salyers A. A. Use of suppressor analysis to find genes involved in the colonization deficiency of a Bacteroides thetaiotaomicron mutant unable to grow on the host-derived mucopolysaccharides chondroitin sulfate and heparin. Appl Environ Microbiol. 1995 Feb;61(2):734–740. doi: 10.1128/aem.61.2.734-740.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooper R., Bush K., Principe P. A., Trejo W. H., Wells J. S., Sykes R. B. Two new monobactam antibiotics produced by a Flexibacter sp. I. Taxonomy, fermentation, isolation and biological properties. J Antibiot (Tokyo) 1983 Oct;36(10):1252–1257. doi: 10.7164/antibiotics.36.1252. [DOI] [PubMed] [Google Scholar]
- Dyer D. W., Bilalis G., Michel J. H., Malek R. Conjugal transfer of plasmid and transposon DNA from Escherichia coli into Porphyromonas gingivalis. Biochem Biophys Res Commun. 1992 Jul 31;186(2):1012–1019. doi: 10.1016/0006-291x(92)90847-e. [DOI] [PubMed] [Google Scholar]
- Gherna R., Woese C. R. A partial phylogenetic analysis of the "flavobacter-bacteroides" phylum: basis for taxonomic restructuring. Syst Appl Microbiol. 1992 Dec;15(4):513–521. doi: 10.1016/S0723-2020(11)80110-4. [DOI] [PubMed] [Google Scholar]
- Hanahan D., Jessee J., Bloom F. R. Plasmid transformation of Escherichia coli and other bacteria. Methods Enzymol. 1991;204:63–113. doi: 10.1016/0076-6879(91)04006-a. [DOI] [PubMed] [Google Scholar]
- Hedges R. W., Jacob A. E. Transposition of ampicillin resistance from RP4 to other replicons. Mol Gen Genet. 1974;132(1):31–40. doi: 10.1007/BF00268228. [DOI] [PubMed] [Google Scholar]
- Herrero M., de Lorenzo V., Timmis K. N. Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria. J Bacteriol. 1990 Nov;172(11):6557–6567. doi: 10.1128/jb.172.11.6557-6567.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirsch P. R., Beringer J. E. A physical map of pPH1JI and pJB4JI. Plasmid. 1984 Sep;12(2):139–141. doi: 10.1016/0147-619x(84)90059-3. [DOI] [PubMed] [Google Scholar]
- Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
- Hwa V., Shoemaker N. B., Salyers A. A. Direct repeats flanking the Bacteroides transposon Tn4351 are insertion sequence elements. J Bacteriol. 1988 Jan;170(1):449–451. doi: 10.1128/jb.170.1.449-451.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katayama N., Fukusumi S., Funabashi Y., Iwahi T., Ono H. TAN-1057 A-D, new antibiotics with potent antibacterial activity against methicillin-resistant Staphylococcus aureus. Taxonomy, fermentation and biological activity. J Antibiot (Tokyo) 1993 Apr;46(4):606–613. doi: 10.7164/antibiotics.46.606. [DOI] [PubMed] [Google Scholar]
- Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
- Li L. Y., Shoemaker N. B., Salyers A. A. Location and characteristics of the transfer region of a Bacteroides conjugative transposon and regulation of transfer genes. J Bacteriol. 1995 Sep;177(17):4992–4999. doi: 10.1128/jb.177.17.4992-4999.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liebert C. A., Hood M. A., Deck F. H., Bishop K., Flaherty D. K. Isolation and characterization of a new Cytophaga species implicated in a work-related lung disease. Appl Environ Microbiol. 1984 Nov;48(5):936–943. doi: 10.1128/aem.48.5.936-943.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mayatepek E., Zilow E., Pohl S. Severe intrauterine infection due to Capnocytophaga ochracea. Biol Neonate. 1991;60(3-4):184–186. doi: 10.1159/000243406. [DOI] [PubMed] [Google Scholar]
- Meyer R. J., Shapiro J. A. Genetic organization of the broad-host-range IncP-1 plasmid R751. J Bacteriol. 1980 Sep;143(3):1362–1373. doi: 10.1128/jb.143.3.1362-1373.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakagawa Y., Yamasato K. Phylogenetic diversity of the genus Cytophaga revealed by 16S rRNA sequencing and menaquinone analysis. J Gen Microbiol. 1993 Jun;139(Pt 6):1155–1161. doi: 10.1099/00221287-139-6-1155. [DOI] [PubMed] [Google Scholar]
- Priefer U. B., Simon R., Pühler A. Extension of the host range of Escherichia coli vectors by incorporation of RSF1010 replication and mobilization functions. J Bacteriol. 1985 Jul;163(1):324–330. doi: 10.1128/jb.163.1.324-330.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salyers A. A., Pajeau M., McCarthy R. E. Importance of mucopolysaccharides as substrates for Bacteroides thetaiotaomicron growing in intestinal tracts of exgermfree mice. Appl Environ Microbiol. 1988 Aug;54(8):1970–1976. doi: 10.1128/aem.54.8.1970-1976.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salyers A. A., Shoemaker N. B., Guthrie E. P. Recent advances in Bacteroides genetics. Crit Rev Microbiol. 1987;14(1):49–71. doi: 10.3109/10408418709104435. [DOI] [PubMed] [Google Scholar]
- Sasakawa C., Yoshikawa M. A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis. Gene. 1987;56(2-3):283–288. doi: 10.1016/0378-1119(87)90145-4. [DOI] [PubMed] [Google Scholar]
- Schoeb T. R., Dybvig K., Davidson M. K., Davis J. K. Cultivation of cilia-associated respiratory bacillus in artificial medium and determination of the 16S rRNA gene sequence. J Clin Microbiol. 1993 Oct;31(10):2751–2757. doi: 10.1128/jcm.31.10.2751-2757.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shi W., Köhler T., Zusman D. R. Chemotaxis plays a role in the social behaviour of Myxococcus xanthus. Mol Microbiol. 1993 Aug;9(3):601–611. doi: 10.1111/j.1365-2958.1993.tb01720.x. [DOI] [PubMed] [Google Scholar]
- Shoemaker N. B., Anderson K. L., Smithson S. L., Wang G. R., Salyers A. A. Conjugal transfer of a shuttle vector from the human colonic anaerobe Bacteroides uniformis to the ruminal anaerobe Prevotella (Bacteroides) ruminicola B(1)4. Appl Environ Microbiol. 1991 Aug;57(8):2114–2120. doi: 10.1128/aem.57.8.2114-2120.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Barber R. D., Salyers A. A. Cloning and characterization of a Bacteroides conjugal tetracycline-erythromycin resistance element by using a shuttle cosmid vector. J Bacteriol. 1989 Mar;171(3):1294–1302. doi: 10.1128/jb.171.3.1294-1302.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Getty C., Gardner J. F., Salyers A. A. Tn4351 transposes in Bacteroides spp. and mediates the integration of plasmid R751 into the Bacteroides chromosome. J Bacteriol. 1986 Mar;165(3):929–936. doi: 10.1128/jb.165.3.929-936.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Guthrie E. P., Salyers A. A., Gardner J. F. Evidence that the clindamycin-erythromycin resistance gene of Bacteroides plasmid pBF4 is on a transposable element. J Bacteriol. 1985 May;162(2):626–632. doi: 10.1128/jb.162.2.626-632.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tancula E., Feldhaus M. J., Bedzyk L. A., Salyers A. A. Location and characterization of genes involved in binding of starch to the surface of Bacteroides thetaiotaomicron. J Bacteriol. 1992 Sep;174(17):5609–5616. doi: 10.1128/jb.174.17.5609-5616.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ubben D., Schmitt R. Tn1721 derivatives for transposon mutagenesis, restriction mapping and nucleotide sequence analysis. Gene. 1986;41(2-3):145–152. doi: 10.1016/0378-1119(86)90093-4. [DOI] [PubMed] [Google Scholar]
- de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. J Bacteriol. 1990 Nov;172(11):6568–6572. doi: 10.1128/jb.172.11.6568-6572.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]