Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101291488 Publication Model: eCollection Cited Medium: Internet ISSN: 1935-2735 (Electronic) Linking ISSN: 19352727 NLM ISO Abbreviation: PLoS Negl Trop Dis Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Relapsing Fever*/microbiology ; Borrelia*/genetics ; Borrelia*/classification; Animals ; Mice ; Female ; Genetic Complementation Test ; Green Fluorescent Proteins/genetics ; Humans

Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Europe) and New World RF Borrelia (found in the Americas). While RF is a rarely reported disease in the Americas, RF is prevalent in endemic parts of Africa. Despite phylogenetic differences between Old World and New World RF Borrelia and higher incidence of disease associated with Old World RF spirochete infection, genetic manipulation has only been described in New World RF bacteria. Herein, we report the generation of genetic tools for use in the Old World RF spirochete, Borrelia duttonii. We describe methods for transformation and establish shuttle vector- and integration-based approaches for genetic complementation, creating green fluorescent protein (gfp)-expressing B. duttonii strains as a proof of principle. Allelic exchange mutagenesis was also used to inactivate a homolog of the Borrelia burgdorferi p66 gene, which encodes an important virulence factor, in B. duttonii and demonstrate that this mutant was attenuated in a murine model of RF. Finally, the B. duttonii p66 mutant was complemented using shuttle vector- and cis integration-based approaches. As expected, complemented p66 mutant strains were fully infectious, confirming that P66 is required for optimal mammalian infection. The genetic tools and techniques reported herein represent an important advancement in the study of RF Borrelia that allows for future characterization of virulence determinants and colonization factors important for the enzootic cycle of Old World RF spirochetes.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Jackson-Litteken et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

J Infect Dis. 2002 Sep 15;186(6):782-91. (PMID: 12198612)
Clin Infect Dis. 1998 Jan;26(1):122-31. (PMID: 9455520)
Infect Immun. 2004 Dec;72(12):7147-54. (PMID: 15557639)
J Mol Microbiol Biotechnol. 2003;6(1):29-40. (PMID: 14593251)
PLoS Negl Trop Dis. 2021 Apr 22;15(4):e0009184. (PMID: 33886571)
Methods Mol Biol. 2008;431:69-84. (PMID: 18287748)
PLoS One. 2016 May 19;11(5):e0155798. (PMID: 27195796)
J Bacteriol. 2003 Nov;185(22):6723-7. (PMID: 14594849)
Infect Immun. 2014 Aug;82(8):3324-32. (PMID: 24866803)
Ann Soc Belg Med Trop. 1984;64(4):335-64. (PMID: 6397148)
Vet Sci. 2016;3(3):. (PMID: 28959690)
J Bacteriol. 2011 Mar;193(5):1161-71. (PMID: 21193609)
Emerg Infect Dis. 2011 May;17(5):883-5. (PMID: 21529402)
PLoS Pathog. 2008 Jun 20;4(6):e1000090. (PMID: 18566656)
Clin Lab Med. 2015 Dec;35(4):847-65. (PMID: 26593261)
Mol Microbiol. 2007 Feb;63(3):694-710. (PMID: 17181780)
Mol Microbiol. 1993 May;8(5):967-80. (PMID: 8102774)
Infect Immun. 1997 Sep;65(9):3654-61. (PMID: 9284133)
Tex Med. 1995 May;91(5):56-9. (PMID: 7778052)
J Bacteriol. 2008 Dec;190(24):7885-91. (PMID: 18849429)
Appl Environ Microbiol. 2007 Mar;73(5):1501-13. (PMID: 17220265)
Emerg Infect Dis. 2007 Jan;13(1):117-23. (PMID: 17370524)
PLoS Negl Trop Dis. 2012;6(11):e1924. (PMID: 23209863)
Lancet. 2006 Jul 1;368(9529):37-43. (PMID: 16815378)
Mol Microbiol. 2006 Mar;59(5):1473-84. (PMID: 16468989)
Infect Immun. 2002 Sep;70(9):4798-804. (PMID: 12183522)
Infect Dis Clin North Am. 2008 Sep;22(3):449-68, viii. (PMID: 18755384)
Emerg Infect Dis. 2015 Jun;21(6):1078-80. (PMID: 25988268)
Infect Immun. 1997 Jan;65(1):285-92. (PMID: 8975925)
Clin Microbiol Infect. 2009 May;15(5):407-14. (PMID: 19489923)
Microb Pathog. 2006 Jul;41(1):43-7. (PMID: 16723206)
PLoS Pathog. 2015 Dec 18;11(12):e1005333. (PMID: 26684456)
PLoS Negl Trop Dis. 2013 Oct 31;7(10):e2514. (PMID: 24205425)
Ticks Tick Borne Dis. 2022 Nov;13(6):102052. (PMID: 36223678)
Clin Infect Dis. 1997 Jul;25(1):139-44. (PMID: 9243047)
Acta Obstet Gynecol Scand. 1997 Oct;76(9):834-8. (PMID: 9351408)
PLoS Genet. 2008 Sep 12;4(9):e1000185. (PMID: 18787695)
Gene. 2003 Jan 16;303:131-7. (PMID: 12559574)
Nat Immunol. 2010 Apr;11(4):295-302. (PMID: 20228796)
PLoS Pathog. 2014 Apr 03;10(4):e1004056. (PMID: 24699793)
PLoS Negl Trop Dis. 2021 Mar 11;15(3):e0008564. (PMID: 33705384)
Biochim Biophys Acta. 2010 Jun;1798(6):1197-203. (PMID: 20188698)
Clin Microbiol Infect. 2009 May;15(5):400-6. (PMID: 19489922)
Mol Microbiol. 2001 Feb;39(3):714-21. (PMID: 11169111)
Mol Microbiol. 2003 Jun;48(6):1665-77. (PMID: 12791146)
J Clin Invest. 2009 Dec;119(12):3652-65. (PMID: 19920352)
Infect Immun. 2011 Sep;79(9):3510-7. (PMID: 21708994)
Plasmid. 2012 Jul;68(1):1-12. (PMID: 22289894)
J Clin Microbiol. 2010 Jul;48(7):2484-9. (PMID: 20463158)
Methods Mol Biol. 2018;1690:183-200. (PMID: 29032546)
Appl Environ Microbiol. 2018 Nov 30;84(24):. (PMID: 30315081)
J Bacteriol. 2011 Jul;193(13):3241-5. (PMID: 21551306)
Emerg Infect Dis. 2005 Mar;11(3):474-5. (PMID: 15757570)
Mol Microbiol. 2012 Sep;85(6):1105-18. (PMID: 22758390)
J Bacteriol. 2003 Feb;185(4):1346-56. (PMID: 12562805)
Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4850-5. (PMID: 10781091)
Plasmid. 2005 Jan;53(1):1-13. (PMID: 15631949)
J Bacteriol. 1997 Nov;179(21):6837-42. (PMID: 9352937)
Curr Protoc Microbiol. 2011 Feb;Chapter 12:Unit 12C.4. (PMID: 21400675)
J Bacteriol. 2008 Nov;190(21):7035-42. (PMID: 18757545)
J Exp Med. 1994 Feb 1;179(2):631-42. (PMID: 8294872)
Front Cell Infect Microbiol. 2018 Apr 04;8:98. (PMID: 29670860)
Appl Environ Microbiol. 2010 Nov;76(22):7407-12. (PMID: 20851957)
J Exp Med. 1982 Nov 1;156(5):1297-311. (PMID: 7130900)
Infect Immun. 2004 Oct;72(10):5938-46. (PMID: 15385497)
Curr Issues Mol Biol. 2021;42:307-332. (PMID: 33300496)
Proc Natl Acad Sci U S A. 2005 May 10;102(19):6972-7. (PMID: 15860579)
Infect Immun. 2015 Aug;83(8):3184-94. (PMID: 26015482)
Infect Immun. 2001 May;69(5):3455-9. (PMID: 11292775)
Appl Environ Microbiol. 2017 Feb 15;83(5):. (PMID: 27986725)
Antonie Van Leeuwenhoek. 2014 Jun;105(6):1049-72. (PMID: 24744012)
PLoS One. 2013 Aug 29;8(8):e72550. (PMID: 24009690)
Mol Microbiol. 1999 Dec;34(5):926-40. (PMID: 10594819)
Mol Microbiol. 2002 Jan;43(2):281-95. (PMID: 11985709)
Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):13142-7. (PMID: 12271135)
Microbiology (Reading). 2003 Jul;149(Pt 7):1819-1828. (PMID: 12855733)
J Appl Microbiol. 2010 Apr;108(4):1115-22. (PMID: 19886891)
Nat Rev Microbiol. 2004 Jan;2(1):67-72. (PMID: 15035010)
J Bacteriol. 1992 Aug;174(16):5251-7. (PMID: 1644750)
Front Cell Infect Microbiol. 2019 Aug 13;9:290. (PMID: 31456953)
Mol Microbiol. 2007 Jun;64(5):1358-74. (PMID: 17542926)
Infect Immun. 2002 Apr;70(4):2139-50. (PMID: 11895980)
J Med Entomol. 2010 Nov;47(6):1164-72. (PMID: 21175068)
Front Cell Infect Microbiol. 2023 Jan 20;13:1112952. (PMID: 36743301)
Int J Syst Bacteriol. 1995 Oct;45(4):804-10. (PMID: 7547303)
Infect Immun. 2021 May 17;89(6):. (PMID: 33846120)
Genetics. 1992 Oct;132(2):311-24. (PMID: 1427031)
Mol Microbiol. 2021 Feb;115(2):191-207. (PMID: 32979869)
Yale J Biol Med. 1984 Jul-Aug;57(4):521-5. (PMID: 6393604)
PLoS Pathog. 2015 Sep 15;11(9):e1005160. (PMID: 26371761)
BMC Microbiol. 2008 May 28;8:82. (PMID: 18507835)
Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7301-6. (PMID: 12748384)
Curr Top Microbiol Immunol. 2018;415:113-140. (PMID: 28918538)
Rev Infect Dis. 1988 Jul-Aug;10 Suppl 2:S274-6. (PMID: 3055197)
Proc Natl Acad Sci U S A. 1989 Apr;86(7):2172-5. (PMID: 2648393)
Gene. 1997 Jun 3;191(2):149-53. (PMID: 9218713)
J Bacteriol. 2000 May;182(9):2445-52. (PMID: 10762244)
Science. 1982 Jun 18;216(4552):1317-9. (PMID: 7043737)
FEMS Microbiol Lett. 2007 Jan;266(2):241-9. (PMID: 17233736)
Nat Rev Dis Primers. 2016 Dec 15;2:16090. (PMID: 27976670)
PLoS Negl Trop Dis. 2014 Apr 03;8(4):e2767. (PMID: 24699275)
Front Public Health. 2014 May 14;2:43. (PMID: 25229054)
Mol Microbiol. 2007 Oct;66(1):262-76. (PMID: 17784908)
Cell Microbiol. 2015 Jul;17(7):1021-36. (PMID: 25604835)

P20 GM103625 United States GM NIGMS NIH HHS; P30 GM145393 United States GM NIGMS NIH HHS; R03 AI151432 United States AI NIAID NIH HHS

147336-22-9 (Green Fluorescent Proteins)

Date Created: 20240722 Date Completed: 20240801 Latest Revision: 20240803

20240803

PMC11293673

10.1371/journal.pntd.0012348

39038047