The Way Forward: Improving Genetic Systems

Ulrike G. Munderloh; Roderick F. Felsheim; Nicole Y. Burkhardt; Michael J. Herron; Adela S. Oliva Chávez; Curtis M. Nelson; Timothy J. Kurtti

doi:10.1128/9781555817336.ch14

Chapter 14 : The Way Forward: Improving Genetic Systems

Authors: Ulrike G. Munderloh¹, Roderick F. Felsheim¹, Nicole Y. Burkhardt¹, Michael J. Herron¹, Adela S. Oliva Chávez¹, Curtis M. Nelson¹, Timothy J. Kurtti¹

VIEW AFFILIATIONS HIDE AFFILIATIONS

Affiliations: 1: Department of Entomology, University of Minnesota, St. Paul, MN 55108

Content Type: Monograph

Publication Year: 2012

Category: Bacterial Pathogenesis

Book DOI: 10.1128/9781555817336

Chapter DOI: 10.1128/9781555817336.ch14

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Ebook: Choose a downloadable PDF or ePub file. Chapter is a downloadable PDF file. File must be downloaded within 48 hours of purchase

Buy this Chapter

Preview this chapter:

The Way Forward: Improving Genetic Systems, Page 1 of 2

< Previous page | Next page > /docserver/preview/fulltext/10.1128/9781555817336/9781555816773_Chap14-1.gif /docserver/preview/fulltext/10.1128/9781555817336/9781555816773_Chap14-2.gif

Abstract:

This chapter reviews the research that has led to the first successes in genetic transformation of arthropod-borne bacteria belonging to the order Rickettsiales in the Alphaproteobacteria subdivision. Seminal results provided the very first evidence that direct genetic manipulation of rickettsiae was achievable, and that the bacteria were able to maintain and express foreign genetic sequences inserted via allelic exchange/homologous recombination under control of rickettsial or Escherichia coli promoters. This work set the stage for all subsequent research efforts aspiring to manipulate and analyze rickettsiae in a manner that is nearly commonplace in extracellular bacteria. The Himar1 transposase allele A7 has been successfully used for mutagenesis of A.phagocytophilum and fever groups of rickettsial tick symbionts have been spotted using fluorescent markers and antibiotic resistance. Selection of mutants using growth inhibitors is an indispensible strategy to recover mutants from the background of nontransformed bacteria. The most common strategy is incorporation of an antibiotic resistance gene in the transformation cassette. When working with human or animal pathogens, this is a sensitive issue, as introduction of resistance to antibiotics used to treat disease induced by the pathogen to be transformed is not encouraged. This poses a dilemma, as the most effective selection is likely achieved by using the clinically most effective antibiotics. The authors suggests that shuttle vectors would be useful for testing the function of genes that are naturally defective in certain Rickettsia spp., in complementation assays, overexpression of native or foreign genes, testing gene regulation, and other applications carried out in E.coli.

Citation: Munderloh U, Felsheim R, Burkhardt N, Herron M, Oliva Chávez A, Nelson C, Kurtti T. 2012. The Way Forward: Improving Genetic Systems, p 416-432. In Palmer G, Azad A (ed), Intracellular Pathogens II: Rickettsiales. ASM Press, Washington, DC. doi: 10.1128/9781555817336.ch14

Key Concept Ranking

Mobile Genetic Elements: 0.65582335
Outer Membrane Proteins: 0.473067
Fatty Acid Biosynthesis: 0.43347782

0.65582335

Highlighted Text: Show | Hide

Full text loading...

Figures

The Way Forward: Improving Genetic Systems

[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-1,webId=/content/author/10.1128/9781555817336.chap14-1,properties={foaf_givenname=Ulrike G., foaf_name=Ulrike G. Munderloh, foaf_surname=Munderloh, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-2,webId=/content/author/10.1128/9781555817336.chap14-2,properties={foaf_givenname=Roderick F., foaf_name=Roderick F. Felsheim, foaf_surname=Felsheim, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-3,webId=/content/author/10.1128/9781555817336.chap14-3,properties={foaf_givenname=Nicole Y., foaf_name=Nicole Y. Burkhardt, foaf_surname=Burkhardt, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-4,webId=/content/author/10.1128/9781555817336.chap14-4,properties={foaf_givenname=Michael J., foaf_name=Michael J. Herron, foaf_surname=Herron, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-5,webId=/content/author/10.1128/9781555817336.chap14-5,properties={foaf_givenname=Adela S., foaf_name=Adela S. Oliva Chávez, foaf_surname=Oliva Chávez, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-6,webId=/content/author/10.1128/9781555817336.chap14-6,properties={foaf_givenname=Curtis M., foaf_name=Curtis M. Nelson, foaf_surname=Nelson, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817336.chap14-7,webId=/content/author/10.1128/9781555817336.chap14-7,properties={foaf_givenname=Timothy J., foaf_name=Timothy J. Kurtti, foaf_surname=Kurtti, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555817336.chap14-aff1]]}]]

/content/10.1128/9781555817336.chap14.fig14-1

FIGURE 1

General features of plasmid constructs being developed for use in Rickettsiales genetics. (A) Construct for transposon-mediated mutagenesis of Anaplasma. Depicted is a cis-plasmid construct: both transposase (Himar1) and a transposon containing a bar (spectinomycin resistance) gene and GFPuv are encoded on a single plasmid (pCis GFPuv Himar1 A7) in order to improve efficiency. In this construct two A. marginale Am tr promoters are used to drive expression of Himar1 and the reporter and resistance genes. (B) Confocal fluorescent microscopic image of GFPuv-expressing A. phagocytophilum infecting a rhesus cell (RF/6A) expressing DsRed2, a red fluorescent protein. (C) Shuttle vector for transformation of rickettsiae. Depicted is a generalized example of the essential elements of shuttle vectors based on pRAM18 from R. amblyommii. A fragment of pRAM18 containing parA and the DnaA-likeprotein gene was ligated into an E. coli cloning vector containing genes for rifampin resistance and GFPuv. (D) Fluorescent microscopic image of pRAM18dRGA-transformed R. bellii strain 369 in an I. scapularis cell (cell line ISE6). Bars (B and D), 5 µm. doi:10.1128/9781555817336.ch14.f1

10.1128/9781555817336/fig14-1_thmb.gif

10.1128/9781555817336/fig14-1.gif

FIGURE 1

References

/content/book/10.1128/9781555817336.chap14

1. Andersson, J. O.,, and S. E. G. Andersson. 1999. Genome degradation is an ongoing process in Rickettsia. Mol. Biol. Evol. 16:1178–1191. PubMed CrossRef

2. Arora, S. K.,, M. Bangera,, S. Lory,, and R. Ramphal. 2001. A genomic island in Pseudomonas aeruginosa carries the determinants of flagellin glycosylation. Proc. Natl. Acad. Sci. USA 98:9342–9347. PubMed CrossRef

3. Baldridge, G. D.,, N. Y. Burkhardt,, R. E. Felsheim,, T. J. Kurtti,, and U. G. Munderloh. 2007a. Transposon insertion reveals pRM, a plasmid of Rickettsia monacensis. Appl. Environ. Microbiol. 73:4984–4995. PubMed CrossRef

4. Baldridge, G. D.,, N. Y. Burkhardt,, R. F. Felsheim,, T. J. Kurtti,, and U. G. Munderloh. 2008. Plasmids of the pRM/pRF family occur in diverse Rickettsia species. Appl. Environ. Microbiol. 74:645–652. PubMed CrossRef

5. Baldridge, G. D.,, N. Burkhardt,, M. J. Herron,, T. J. Kurtti,, and U. G. Munderloh. 2005. Analysis of fluorescent protein expression in transformants of Rickettsia monacensis, an obligate intracellular tick symbiont. Appl. Environ. Microbiol. 71:2095–2105. PubMed CrossRef

6. Baldridge, G. D.,, N. Y. Burkhardt,, M. B. Labruna,, R. C. Pacheco,, C. D. Paddock,, P. C. Williamson,, P. M. Billingsley,, R. F. Felsheim,, T. J. Kurtti,, and U. G. Munderloh. 2010a. Wide dispersal and possible multiple origins of low-copy-number plasmids in Rickettsia species associated with blood-feeding arthropods. Appl. Environ. Microbiol. 76:1718–1731. PubMed CrossRef

7. Baldridge, G. D.,, N. Y. Burkhardt,, A. S. Oliva,, T. J. Kurtti,, and U. G. Munderloh. 2010b. Rickettsial ompB promoter regulated expression of GFPuv in transformed Rickettsia montanensis. PLoS One 5:e8965. PubMed CrossRef

8. Baldridge, G. D.,, T. J. Kurtti,, N. Burkhardt,, A. S. Baldridge,, C. M. Nelson,, A. S. Oliva,, and U. G. Munderloh. 2007b. Infection of Ixodes scapularis ticks with Rickettsia monacensis expressing green fluorescent protein: a model system. J. Invertebr. Pathol. 94:163–174. PubMed CrossRef

9. Barbet, A. F.,, J. T. Agnes,, A. L. Moreland,, A. M. Lundgren,, A. R. Alleman,, S. M. Noh,, K. A. Brayton,, U. G. Munderloh,, and G. H. Palmer. 2005. Identification of functional promoters in the msp2 expression loci of Anaplasma marginale and Anaplasma phagocytophilum. Gene 353:89–97. PubMed CrossRef

10. Bassler, L. 1999. How bacteria talk to each other: regulation of gene expression by quorum sensing. Curr. Opin. Microbiol. 2:582–587. PubMed CrossRef

11. Bechah, Y.,, K. El Karkouri,, O. Mediannikov,, Q. Leroy,, N. Pelletier,, C. Robert,, C. Médigue,, J. L. Mege,, and D. Raoult. 2010. Genomic, proteomic, and transcriptomic analysis of virulent and avirulent Rickettsia prowazekii reveals its adaptive mutation capabilities. Genome Res. 20:655–663. PubMed CrossRef

12. Bertram, R.,, and W. Hillen. 2008. The application of Tet repressor in prokaryotic gene regulation and expression. Microb. Biotechnol. 1:2–16. PubMed CrossRef

13. Blanc, G.,, H. Ogata,, C. Robert,, S. Audic,, J. M.. Claverie, and D. Raoult. 2007. Lateral gene transfer between obligate intracellular bacteria: evidence from the Rickettsia massiliae genome. Genome Res. 17:1657–1664. PubMed CrossRef

14. Bouet, J.-Y.,, K. Nordstrom,, and D. Lane. 2007. Plasmid partition and incompatibility—the focus shifts. Mol. Microbiol. 65:1405–1414. PubMed CrossRef

15. Boussau, B.,, E. O. Karlberg,, A. C. Frank,, B. A. Legault,, and S. G. E. Andersson. 2004. Computational inference of scenarios for α-proteobacterial genome evolution. Proc. Natl. Acad. Sci. USA 101:9722–9727. PubMed CrossRef

16. Brayton, K. A.,, L. S. Kappmeyer,, D. R. Herndon,, M. J. Dark,, D. L. Tibbals,, G. H. Palmer,, T. C. McGuire,, and D. P. Knowles, Jr. 2005. Complete genome sequencing of Anaplasma marginale reveals that the surface is skewed to two superfamilies of outer membrane proteins. Proc. Natl. Acad. Sci. USA 102:844–849. PubMed CrossRef

17. Brouqui, P.,, J. R. Harle,, J. Delmont,, C. Frances,, P. J. Weiller,, and D. Raoult. 1997. African tick-bite fever. An imported spotless rickettsiosis. Arch. Intern. Med. 157:119–124. PubMed CrossRef

18. Burkhardt, N. Y.,, G. D. Baldridge,, P. C. Williamson,, P. M. Billingsley,, R. F. Felsheim,, T. J. Kurtti,, and U. G. Munderloh. 2010. Development of shuttle vectors for transformation of diverse Rickettsia species, PLoS One 6:e29511. PubMed CrossRef

19. Cardwell, M. M.,, and J. J. Martinez. 2009. The Sca2 autotransporter protein from Rickettsia conorii is sufficient to mediate adherence to and invasion of cultured mammalian cells. Infect. Immun. 77:5272–5280. PubMed CrossRef

20. Cevallos, M. A.,, R. Cervantes-Rivera,, and R. M.. Gutiérrez-Ríos. 2008. The repABC plasmid family. Plasmid 60:19–37. PubMed CrossRef

21. Chan, Y. G.,, M. M. Cardwell,, T. M. Hermanas,, T. Uchiyama,, and J. J. Martinez. 2009. Rickettsial outer-membrane protein B (rOmpB) mediates bacterial invasion through Ku70 in an actin, c-Cbl, clathrin and caveolin 2-dependent manner. Cell. Microbiol. 11:629–644. PubMed CrossRef

22. Clark, T. R.,, D. W. Ellison,, B. Kleba,, and T. Hackstadt. 2011. Complementation of Rickettsia rickettsii RelA/SpoT restores a non-lytic plaque phenotype. Infect. Immun. 79:1631–1637. PubMed CrossRef

23. Collins, N. E.,, J. Liebenberg,, E. P. de Villiers,, K. A. Brayton,, E. Louw,, A. Pretorius,, F. E. Faber,, H. van Heerden,, A. Josemans,, M. van Kleef,, H. C. Steyn,, M. F. van Strijp,, E. Zweygarth,, F. Jongejan,, J. C. Maillard,, D. Berthier,, M. Botha,, F. Joubert,, C. H. Corton,, N. R. Thomson,, M. T. Allsopp,, and B. A. Allsopp. 2005. The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. Proc. Natl. Acad. Sci. USA 102:838–843. PubMed CrossRef

24. Crameri, A.,, E. A. Whitehorn,, E. Tate,, and W. P.. Stemmer. 1996. Improved green fluorescent protein by molecular evolution using DNA shuffling. Nat. Biotechnol. 14:315–319. PubMed CrossRef

25. de la Fuente, J.,, J. C. Garcia-Garcia,, E. F. Blouin,, and K. M. Kocan. 2001. Differential adhesion of major surface proteins 1a and 1b of the ehrlichial cattle pathogen Anaplasma marginale to bovine erythrocytes and tick cells. Int. J. Parasitol. 31:145–153. PubMed CrossRef

26. de Sousa, R.,, L. Duque,, J. Poças,, J. Torgal,, F. Bacellar,, J. P. Olano,, and D. H. Walker. 2008. Lymphangitis in a Portuguese patient infected with Rickettsia sibirica. Emerg. Infect. Dis. 14:529–531. PubMed CrossRef

27. Donovan, B. J.,, D. J. Weber,, J. C. Rublein,, and R. H. Raasch. 2002. Treatment of tick-borne diseases. Ann. Pharmacother. 36:1590–1597. PubMed CrossRef

28. Doré, M.,, R. J. Korthuis,, D. N. Granger,, M. L. Entman,, and C. W. Smith. 1993. P-selectin mediates spontaneous leukocyte rolling in vivo. Blood 82:1308–1316. PubMed

29. Driskell, L. O.,, X.-J. Yu,, L. Zhang,, Y. Liu,, V. L. Popov,, D. H. Walker,, A. M. Tucker,, and D. O. Wood. 2009. Directed mutagenesis of the Rickettsia prowazekii pld gene encoding phospholipase D. Infect. Immun. 77:3244–3248. PubMed CrossRef

30. Dunning Hotopp, J. C.,, M. Lin,, R. Madupu,, J. Crabtree,, S. V. Angiuoli,, J. Eisen,, R. Seshadri,, Q. Ren,, M. Wu,, T. R. Utterback,, S. Smith,, M. Lewis,, H. Khouri,, C. Zhang,, H. Niu,, Q. Lin,, N. Ohashi,, N. Zhi,, W. Nelson,, L. M. Brinkac,, R. J. Dodson,, M. J. Rosovitz,, J. Sundaram,, S. C. Daugherty,, T. Davidsen,, A. S. Durkin,, M. Gwinn,, D. H. Haft,, J. D. Selengut,, S. A. Sullivan,, N. Zafar,, L. Zhou,, F. Benahmed,, H. Forberger,, R. Halpin,, S. Mulligan,, J. Robinson,, O. White,, Y. Rikihisa,, and H. Tettelin. 2006. Comparative genomics of emerging human ehrlichiosis agents. PLoS Genet. 2:e21. PubMed CrossRef

31. Ellison, D. W.,, T. R. Clark,, D. E. Sturdevant,, K. Virtaneva,, S. F. Porcella,, and T. Hackstadt. 2008. Genomic comparison of virulent Rickettsia rickettsii Sheila Smith and avirulent Rickettsia rickettsii Iowa. Infect. Immun. 76:542–550. PubMed CrossRef

32. Ettema, T. J. G.,, and S. G. E. Andersson. 2009. The α-proteobacteria: the Darwin finches of the bacterial world. Biol. Lett. 5:429–432. PubMed CrossRef

33. Felsheim, R. F.,, M. J. Herron,, C., and U. G. Munderloh. 2007. Selection of Anaplasma phagocytophilum transformants with an herbicide resistance system, abstr. 3. 21st Meet. Am. Soc. Rickettsiol., Colorado Springs, CO, 8 to 11 September 2007.

34. Felsheim, R. F.,, M. J. Herron,, C. M. Nelson,, N. Y. Burkhardt,, A. F. Barbet,, T. J. Kurtti,, and U. G. Munderloh. 2006. Transformation of Anaplasma phagocytophilum. BMC Biotechnol. 6:42. PubMed CrossRef

35. Felsheim, R. F.,, T. J. Kurtti,, and U. G. Munderloh. 2009. Genome sequence of the endosymbiont Rickettsia peacockii and comparison with virulent Rickettsia rickettsii: identification of virulence factors. PLoS One 4:e8361. PubMed CrossRef

36. Felsheim, R. F.,, A. S. Oliva Chávez,, G. H. Palmer,, L. Crosby,, A. F. Barbet,, T. J. Kurtti,, and U. G. Munderloh. 2010. Transformation of Anaplasma marginale. Vet. Parasitol. 167:167–174. PubMed CrossRef

37. Fournier, P. E.,, K. El Karkouri,, Q. Leroy,, C. Robert,, B. Giumelli,, P. Renesto,, C. Socolovschi,, P. Parola,, S. Audic,, and D. Raoult. 2009. Analysis of the Rickettsia africae genome reveals that virulence acquisition in Rickettsia species may be explained by genome reduction. BMC Genomics 10:166. PubMed CrossRef

38. Fournier, P. E.,, F. Gouriet,, P. Brouqui,, F. Lucht,, and D. Raoult. 2005. Lymphangitis-associated rickettsiosis, a new rickettsiosis caused by Rickettsia sibirica mongolotimonae: seven new cases and review of the literature. Clin. Infect. Dis. 40:1435–1444. PubMed CrossRef

39. Funnell, B. 2005. Partition-mediated plasmid pairing. Plasmid 53:119–125. PubMed CrossRef

40. Fuxelius, H. H.,, A. C. Darby,, N. H. Cho,, and S. G. E. Andersson. 2008. Visualization of pseudogenes in intracellular bacteria reveals the different tracks to gene destruction. Genome Biol. 9:R42. PubMed CrossRef

41. Fuxelius, H. H.,, A. Darby,, C. K. Min,, N. M. Cho,, and S. G. E. Andersson. 2007. The genomic and metabolic diversity of Rickettsia. Res. Microbiol. 158:745–753.

42. Garcia-Garcia, J. C.,, J. de la Fuente,, G. Bell-Eunice,, E. F. Blouin,, and K. M. Kocan. 2004. Glycosylation of Anaplasma marginale major surface protein 1a and its putative role in adhesion to tick cells. Infect. Immun. 72:3022–3030. PubMed CrossRef

43. Gill, H. S.,, and D. Eisenberg. 2001. The crystal structure of phosphinothricin in the active site of glutamine synthetase illuminates the mechanism of enzymatic inhibition. Biochemistry 40:1903–1912. PubMed CrossRef

44. Gillespie, J. J.,, M. S. Beier,, M. S. Rahman,, N. C. Ammerman,, J. M. Shallom,, A. Purkayastha,, B. S. Sobral,, and A. F. Azad. 2007. Plasmids and rickettsial evolution: insight from Rickettsia felis. PLoS One 2:e266. PubMed CrossRef

45. Gillespie, J. J.,, K. Williams,, M. Shukla,, E. E. Snyder,, E. K. Nordberg,, S. M. Ceraul,, C. Dharmanolla,, D. Rainey,, J. Soneja,, J. M. Shallom,, N. D. Vishnubhat,, R. Wattam,, A. Purkayastha,, M. Czar,, O. Crasta,, J. C. Setubal,, A. F. Azad,, and B. S. Sobral. 2008. Rickettsia phylogenomics: unwinding the intricacies of obligate intracellular life. PLoS One 3:e2018. PubMed

46. Grundy, F. J., and T. M. Henkin. 2006. From ribosome to riboswitch: control of gene expression in bacteria by RNA structural rearrangements. Crit. Rev. Biochem. Mol. Biol. 41:329–338. PubMed CrossRef

47. Herron, M. J.,, C. M. Nelson,, J. Larson,, K. R. Snapp,, G. S. Kansas,, and J. L. Goodman. 2000. Intracellular parasitism by the human granulocytic ehrlichiosis bacterium through the P-selectin ligand, PSGL-1. Science 288:1653–1656. PubMed CrossRef

48. Herron, P. R.,, G. Hughes,, G. Chandra,, S. Fielding,, and P. J. Dyson. 2004. Transposon Express, a software application to report the identity of insertions obtained by comprehensive transposon mutagenesis of sequenced genomes: analysis of the preference for in vitro Tn5 transposition into GC-rich DNA. Nucleic Acids Res. 32:e113. PubMed CrossRef

49. Holman, R. C.,, C. D. Paddock,, A. T. Curns,, J. W. Krebs,, J. H. McQuiston,, and J. E. Childs. 2001. Analysis of risk factors for fatal Rocky Mountain spotted fever: evidence for superiority of tetracyclines for treatment. J. Infect. Dis. 184:1437–1444. PubMed CrossRef

50. Kenyon, R. H.,, W. M. Acree,, G. G. Wright,, and F. W. Melchior, Jr. 1972. Preparation of vaccines for Rocky Mountain spotted fever from rickettsiae propagated in cell culture. J. Infect. Dis. 125:146–152. PubMed CrossRef

51. Kurtti, T. J.,, J. A. Simser,, G. D. Baldridge,, A. T. Palmer,, and U. G. Munderloh. 2005. Factors influencing in vitro infectivity and growth of Rickettsia peacockii (Rickettsiales: Rickettsiaceae), an endosymbiont of the Rocky Mountain wood tick, Dermacentor andersoni (Acari, Ixodidae). J. Invertebr. Pathol. 90:177–186. PubMed CrossRef

52. Lampe, D. J.,, B. J. Akerley,, E. J. Rubin,, J. J. Mekalanos,, and H. M. Robertson. 1999. Hyperactive transposase mutants of the Himar1 mariner transposon. Proc. Natl. Acad. Sci. USA 96:11428–11433. PubMed CrossRef

53. Lampe, D. J.,, M. E. Churchill,, and H. M. Robertson. 1996. A purified mariner transposase is sufficient to mediate transposition in vitro. EMBO J. 15:5470–5479. PubMed

54. Lampe, D. J.,, T. E. Grant,, and H. M. Robertson. 1998. Factors affecting transposition of the Himar1 mariner transposon in vitro. Genetics 149:179–187. PubMed

55. Li, H.,, and D. H. Walker. 1998. rOmpA is a critical protein for the adhesion of Rickettsia rickettsii to host cells. Microb. Pathog. 24:289–298. PubMed CrossRef

56. Lin, M.,, T. Kikuchi,, H. M. Brewer,, A. D. Norbeck,, and Y. Rikihisa. 2011. Global proteomic analysis of two tick-borne emerging zoonotic agents: Anaplasma phagocytophilum and Ehrlichia chaffeensis. Front. Microbiol. 2:24. PubMed CrossRef

57. Liu, Z.-M.,, A. M. Tucker,, L. O. Driskell,, and D. O. Wood. 2007. Mariner-based transposon mutagenesis of Rickettsia prowazekii. Appl. Environ. Microbiol. 73:6644–6649. PubMed CrossRef

58. Lutz, K. A.,, J. E. Knapp,, and P. Maliga. 2001. Expression of bar in the plastid genome confers herbicide resistance. Plant Physiol. 125:1585–1590. PubMed CrossRef

59. Mahan, M. J.,, J. M. Slauch,, and J. J. Mekalanos. 1993. Selection of bacterial virulence genes that are specifically induced in host tissues. Science 259:686–688. PubMed CrossRef

60. Martinez, J. J.,, S. Seveau,, E. Veiga,, S. Matsuyama,, and P. Cossart. 2005. Ku70, a component of DNA-dependent protein kinase, is a mammalian receptor for Rickettsia conorii. Cell 123:1013–1023. PubMed CrossRef

61. McClintock, B. 1950. The origin and behavior of mutable loci in maize. Proc. Natl. Acad. Sci. USA 36:344–355. PubMed CrossRef

62. McGarey, D. J.,, A. F. Barbet,, G. H. Palmer,, T. C.. McGuire, and D. R. Allred. 1994. Putative adhesins of Anaplasma marginale: major surface polypeptides 1a and 1b. Infect. Immun. 62:4594–4601. PubMed

63. Moran, J. S.,, and W. C. Levine. 1995. Drugs of choice for the treatment of uncomplicated gonococcal infections. Clin. Infect. Dis. 1:S47–S65. PubMed CrossRef

64. Munderloh, U. G.,, S. D. Jauron,, V. Fingerle,, L. Leitritz,, S. F. Hayes,, J. M. Hautman,, C. M. Nelson,, B. W. Huberty,, T. J. Kurtti,, G. G. Ahlstrand,, B. Greig,, M. A. Mellencamp,, and J. L. Goodman. 1999. Invasion and intracellular development of the human granulocytic ehrlichiosis agent in tick cell culture. J. Clin. Microbiol. 37:2518–2524. PubMed

65. Munderloh, U. G.,, and T. J. Kurtti. 1995. Cellular and molecular interrelationships between ticks and prokaryotic tick-borne pathogens. Annu. Rev. Entomol. 40:221–243. PubMed CrossRef

66. Munderloh, U. G.,, M. J. Lynch,, M. J. Herron,, A. T. Palmer,, T. J. Kurtti,, R. D. Nelson,, and J. L. Goodman. 2004. Infection of endothelial cells with Anaplasma marginale and A. phagocytophilum. Vet. Microbiol. 101:53–64. PubMed CrossRef

67. Nelson, C. M.,, M. J. Herron,, R. F. Felsheim,, B. R. Schloeder,, S. M. Grindle,, A. O. Chavez,, T. J. Kurtti,, and U. G. Munderloh. 2008. Whole genome transcription profiling of Anaplasma phagocytophilum in human and tick host cells by tiling array analysis. BMC Genomics 9:364. PubMed CrossRef

68. Niebylski, M. L.,, M. E. Schrumpf,, W. Burgdorfer,, E. R. Fischer,, K. L. Gage,, and T. G.. Schwan. 1997. Rickettsia peacockii sp. nov., a new species infecting wood ticks, Dermacentor andersoni, in western Montana. Int. J. Syst. Bacteriol. 47:446–452. PubMed CrossRef

69. Ogata, H.,, P. Renesto,, S. Audic,, C. Robert,, G. Blanc,, P. E. Fournier,, H. Parinello,, J. M. Claverie,, and D. Raoult. 2005. The genome sequence of Rickettsia felis identifies the first putative conjugative plasmid in an obligate intracellular parasite. PLoS Biol. 3:e248. PubMed CrossRef

70. Oliva Chávez, A. S.,, R. F. Felsheim,, N. Y. Burkhardt,, T. J. Kurtti,, and U. G. Munderloh. 2010. Identification of genes involved in the infection process of tick (ISE6) and human (HL-60) cells by the tick-borne pathogen Anaplasma phagocytophilum. 42nd Annu. Conf. Soc. Vector Ecol., Raleigh, NC, 26 to 30 September 2010.

71. Purvis, J. J.,, and M. S. Edwards. 2000. Doxycycline use for rickettsial disease in pediatric patients. Pediatr. Infect. Dis. J. 19:871–874. PubMed CrossRef

72. Qin, A.,, A. M. Tucker,, A. Hines,, and D. O. Wood. 2004. Transposon mutagenesis of the obligate intracellular pathogen Rickettsia prowazekii. Appl. Environ. Microbiol. 70:2816–2822. PubMed

73. Rachek, L. I.,, A. Hines,, A. M. Tucker,, H. H. Winkler,, and D. O. Wood. 2000. Transformation of Rickettsia prowazekii to erythromycin resistance encoded by the Escherichia coli ereB gene. J. Bacteriol. 182:3289–3291. PubMed

74. Rachek, L. I.,, A. M. Tucker,, H. H. Winkler,, and D. O. Wood. 1998. Transformation of Rickettsia prowazekii to rifampin resistance. J. Bacteriol. 180:2118–2124. PubMed

75. Ramabu, S. S.,, M. W. Ueti,, K. A. Brayton,, T. V. Baszler,, and G. H. Palmer. 2010. Identification of Anaplasma marginale proteins specifically upregulated during colonization of the tick vector. Infect. Immun. 78:3047–3052. PubMed CrossRef

76. Renesto, P.,, E. Gouin,, and D. Raoult. 2002. Expression of green fluorescent protein in Rickettsia conorii. Microb. Pathog. 33:17–21. PubMed CrossRef

77. Renesto, P.,, L. Samson,. H. Ogata,, S. Azza,, P. Fourquet,, J. P. Gorvel,, R. A. Heinzen,, and D. Raoult. 2006. Identification of two putative rickettsial adhesins by proteomic analysis. Res. Microbiol. 157:605–612. PubMed CrossRef

78. Riley, S. P.,, K. C. Goh,, T. M. Hermanas,, M. M. Cardwell,, Y. G. Chan,, and J. J. Martinez. 2010. The Rickettsia conorii autotransporter protein Sca1 promotes adherence to nonphagocytic mammalian cells. Infect. Immun. 78:1895–1904. PubMed CrossRef

79. Sahni, S. K., and E. Rydkina. 2009. Progress in the functional analysis of rickettsial genes through directed mutagenesis of Rickettsia prowazekii phospholipase D. Future Microbiol. 4:1249–1253. PubMed CrossRef

80. Sakharkar, K. R.,, M. K. Sakharkar,, C. Verma,, and V. T. K. Chow. 2005. Comparative study of overlapping genes in bacteria, with special reference to Rickettsia prowazekii and Rickettsia conorii. Int. J. Syst. Evol. Microbiol. 55:1205–1209. PubMed CrossRef

81. Shaner, N. C.,, P. A. Steinbach,, and R. Y. Tsien. 2005. A guide to choosing fluorescent proteins. Nat. Methods 2:905–909. PubMed CrossRef

82. Shapiro, J. A. 1999. Transposable elements as the key to a 21st century view of evolution. Genetica 107:171–179. PubMed CrossRef

83. Simser, J. A.,, M. S. Rahman,, S. M. Dreher-Lesnick,, and A. F. Azad. 2005. A novel and naturally occurring transposon, ISRpe1 in the Rickettsia peacockii genome disrupting the rickA gene involved in actin-based motility. Mol. Microbiol. 58:71–79. PubMed CrossRef

84. Stary, E.,, R. Gaupp,, S. Lechner,, M. Leibig,, E. Tichy,, M. Kolb,, and R. Bertram. 2010. New architectures for Tet-on and Tet-off regulation in Staphylococcus aureus. Appl. Environ. Microbiol. 76:680–687. PubMed CrossRef

85. Stewart, P. E.,, and P. A. Rosa. 2008. Transposon mutagenesis of the Lyme disease agent Borrelia burgdorferi. Methods Mol. Biol. 431:85–95. PubMed CrossRef

86. Suk, J. E.,, A. Zmorzynska,, A. Hunger,, W. Biederbick,, J. Sasse,, H. Maidhof,, and J. C. Semenza. 2011. Dual-use research and technological diffusion: reconsidering the bioterrorism threat spectrum. PLoS Pathog. 7:e1001253. PubMed CrossRef

87. Troyer, J. M.,, S. Radulovic,, and A. F. Azad. 1999. Green fluorescent protein as a marker in Rickettsia typhi transformation. Infect. Immun. 67:3308–3311. PubMed

88. Uchiyama, T. 2003. Adherence to and invasion of Vero cells by recombinant Escherichia coli expressing the outer membrane protein rOmpB of Rickettsia japonica. Ann. N. Y. Acad. Sci. 990:585–590. PubMed

89. Uchiyama, T.,, H. Kawano,, and Y. Kusuhara. 2006. The major outer membrane protein rOmpB of spotted fever group rickettsiae functions in the rickettsial adherence to and invasion of Vero cells. Microbes Infect. 8:801–809. PubMed CrossRef

90. Vellaiswamy, M.,, M. Kowalczewska,, V. Merhej,, C. Nappez,, R. Vincentelli,, P. Renesto,, and D. Raoult. 2011. Characterization of rickettsial adhesin Adr2 belonging to a new group of adhesins in α-proteobacteria. Microb. Pathog. 50:233–242. PubMed CrossRef

91. Walker, D. H.,, G. A. Valbuena,, and J. P. Olano. 2003. Pathogenic mechanisms of diseases caused by Rickettsia. Ann. N. Y. Acad. Sci. 990:1–11. PubMed CrossRef

92. Wu, Q.,, J. Pei,, C. Turse,, and T. A. Ficht. 2006. Mariner mutagenesis of Brucella melitensis reveals genes with previously uncharacterized roles in virulence and survival. BMC Microbiol. 6:102. PubMed CrossRef

Tables

The Way Forward: Improving Genetic Systems

/content/10.1128/9781555817336.chap14.tab14-1

TABLE 1a

Rickettsiales genetic transformation systems

a Rifampin-resistant mutant gene of R. prowazekii Madrid E.

b E. coli ereB gene.

c E. coli srp promoter.

d R. prowazekii-adapted rifampin resistance arr-2 gene.

e CAT, chloramphenicol acetyltransferase.

10.1128/9781555817336/tab14-1a_thmb.gif

10.1128/9781555817336/tab14-1a.gif

TABLE 1a

Rickettsiales genetic transformation systems

a Rifampin-resistant mutant gene of R. prowazekii Madrid E.

b E. coli ereB gene.

c E. coli srp promoter.

d R. prowazekii-adapted rifampin resistance arr-2 gene.

e CAT, chloramphenicol acetyltransferase.

/content/10.1128/9781555817336.chap14.tab14-1b

TABLE 1b

Rickettsiales genetic transformation systems

a Rifampin-resistant mutant gene of R. prowazekii Madrid E.

b E. coli ereB gene.

c E. coli srp promoter.

d R. prowazekii-adapted rifampin resistance arr-2 gene.

e CAT, chloramphenicol acetyltransferase.

10.1128/9781555817336/tab14-1b_thmb.gif

10.1128/9781555817336/tab14-1b.gif

TABLE 1b

Rickettsiales genetic transformation systems

a Rifampin-resistant mutant gene of R. prowazekii Madrid E.

b E. coli ereB gene.

c E. coli srp promoter.

d R. prowazekii-adapted rifampin resistance arr-2 gene.

e CAT, chloramphenicol acetyltransferase.