{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T19:02:04Z","timestamp":1791658924570,"version":"4.3.4"},"reference-count":30,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5568","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2002,4,26]]},"abstract":"<jats:p>\n                    Transmission by flea bite is a relatively recent adaptation that distinguishes\n                    <jats:italic>Yersinia pestis<\/jats:italic>\n                    , the plague bacillus, from closely related enteric bacteria. Here, a plasmid-encoded phospholipase D (PLD), previously characterized as Yersinia murine toxin (Ymt), was shown to be required for survival of\n                    <jats:italic>Y. pestis<\/jats:italic>\n                    in the midgut of its principal vector, the rat flea\n                    <jats:italic>Xenopsylla cheopis<\/jats:italic>\n                    . Intracellular PLD activity appeared to protect\n                    <jats:italic>Y. pestis<\/jats:italic>\n                    from a cytotoxic digestion product of blood plasma in the flea gut. By enabling colonization of the flea midgut, acquisition of this PLD may have precipitated the transition of\n                    <jats:italic>Y. pestis<\/jats:italic>\n                    to obligate arthropod-borne transmission.\n                  <\/jats:p>","DOI":"10.1126\/science.1069972","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:54:32Z","timestamp":1027749272000},"page":"733-735","source":"Crossref","is-referenced-by-count":264,"title":["Role of Yersinia Murine Toxin in Survival of\n                    <i>Yersinia pestis<\/i>\n                    in the Midgut of the Flea Vector"],"prefix":"10.1126","volume":"296","author":[{"given":"B. Joseph","family":"Hinnebusch","sequence":"first","affiliation":[{"name":"Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy E.","family":"Rudolph","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Cherepanov","sequence":"additional","affiliation":[{"name":"Department of Medical Countermeasures, Swedish Defense Research Agency, S-901 82 Ume\u00e5, and Department of Molecular Biology, Ume\u00e5 University, S-901 87 Ume\u00e5, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jack E.","family":"Dixon","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom G.","family":"Schwan","sequence":"additional","affiliation":[{"name":"Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u212bke","family":"Forsberg","sequence":"additional","affiliation":[{"name":"Department of Medical Countermeasures, Swedish Defense Research Agency, S-901 82 Ume\u00e5, and Department of Molecular Biology, Ume\u00e5 University, S-901 87 Ume\u00e5, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","unstructured":"M. Achtman et al. Proc. Natl. Acad. Sci. U.S.A. 96 14043 (1999).","DOI":"10.1073\/pnas.96.24.14043"},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1128\/CMR.4.3.309","volume":"4","author":"Brubaker R. R.","year":"1991","unstructured":"Brubaker R. R., Clin. Microbiol. Rev. 4, 309 (1991).","journal-title":"Clin. Microbiol. Rev."},{"key":"e_1_3_1_4_2","unstructured":"P. A. Cherepanov et al. Mol. Gen. Mikrobiol. Virusol. 12 19 (1991)."},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","unstructured":"P. Hu et al. J. Bacteriol. 180 5192 (1998).","DOI":"10.1128\/JB.180.19.5192-5202.1998"},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","first-page":"5731","DOI":"10.1128\/IAI.66.12.5731-5742.1998","volume":"66","author":"Lindler L. E.","year":"1998","unstructured":"Lindler L. E., Plano G. V., Burland V., Mayhew G. F., Blattner F. R., Infect. Immun. 66, 5731 (1998).","journal-title":"Infect. Immun."},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","unstructured":"A. E. Rudolph et al. J. Biol. Chem. 274 11824 (1999).","DOI":"10.1074\/jbc.274.17.11824"},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1128\/jb.70.2.158-169.1955","volume":"70","author":"Ajl S. F.","year":"1955","unstructured":"Ajl S. F., Reedal J. S., Durrum E. L., Warren J., J. Bacteriol. 70, 158 (1955).","journal-title":"J. Bacteriol."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1128\/iai.18.1.85-93.1977","volume":"18","author":"Brown S. D.","year":"1977","unstructured":"Brown S. D., Montie T. C., Infect. Immun. 18, 85 (1977).","journal-title":"Infect. Immun."},{"key":"e_1_3_1_10_2","first-page":"51","volume":"19","author":"Sch\u00e4r M.","year":"1956","unstructured":"Sch\u00e4r M., Meyer K. F., Schweiz. Z. Pathol. Bakteriol. 19, 51 (1956).","journal-title":"Schweiz. Z. Pathol. Bakteriol."},{"key":"e_1_3_1_11_2","unstructured":"T. C. Montie S. J. Ajl in Microbial Toxins T. C. Montie S. Kadis S. J. Ajl Eds. (Academic Press New York 1970) vol. 3 pp. 1\u201337."},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(96)30024-8"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560050513"},{"key":"e_1_3_1_14_2","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1038\/6716","volume":"6","author":"Stuckey J. A.","year":"1999","unstructured":"Stuckey J. A., Dixon J. E., Nature Struct. Biol. 6, 278 (1999).","journal-title":"Nature Struct. Biol."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","unstructured":"J. Hinnebusch et al. Int. J. Med. Microbiol. 290 483 (2000).","DOI":"10.1016\/S1438-4221(00)80070-3"},{"key":"e_1_3_1_16_2","first-page":"321","volume":"13","author":"Du Y.","year":"1995","unstructured":"Du Y., Galyov E., Forsberg \u00c5., Contrib. Microbiol. Immunol. 13, 321 (1995).","journal-title":"Contrib. Microbiol. Immunol."},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","unstructured":"I. G. Drozdov et al. J. Med. Microbiol. 42 264 (1995).","DOI":"10.1099\/00222615-42-4-264"},{"key":"e_1_3_1_18_2","unstructured":"We infected X. cheopis fleas by allowing them to feed on heparinized mouse blood containing \u223c5.0 \u00d7 10 8 bacteria per milliliter using a membrane feeder apparatus (19 26). In three replicate experiments with each bacterial strain equal numbers of male and female fleas that took the single infectious blood meal were kept at 21\u00b0C in an atmosphere of 75% relative humidity and subsequently fed twice weekly on uninfected mice. Fleas were monitored for 4 weeks for mortality and proventricular blockage as described (19). Infection rates and levels were determined by colony-forming unit counts (19) from 20 individually triturated female fleas collected at 1 hour 1 day 7 days and 28 days after the infectious blood meal. A mixed infection with Y. pestis KIM6+ and with Y. pestis KIM6+ ymt H188N that had been transformed with pGFP (Clontech) was established by feeding fleas on blood containing 4.0 \u00d7 10 8 of both bacteria per millileter. The pGFP plasmid carries the gfp gene and is stable in Y. pestis even in the absence of selective pressure (19). One to 3 weeks after the infectious blood meal dissected digestive tracts of 64 fleas that contained typical bacterial aggregates were examined by light and fluorescence microscopy. To test the effect of exogenous Ymt in the flea gut we infected fleas using mouse blood containing 140 \u03bcg of Ymt protein per millileter (6 13). To test the effect of blood components we infected fleas with filtered mouse plasma (Millipore Millex-GS 0.45-\u03bcm pore diameter) or artificial plasma [phosphate-buffered saline (PBS pH 7.4) containing 7% bovine serum albumin (BSA) 6 mM glucose 12 mM sodium bicarbonate 10 mM MgCl 2 2.5 mM CaCl 2 and 1 mM citric acid] with or without an equal volume of washed mouse red blood cells."},{"key":"e_1_3_1_19_2","unstructured":"A. W. Bacot C. J. Martin J. Hyg. (suppl. 3) 423 (1914)."},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.273.5273.367"},{"key":"e_1_3_1_21_2","unstructured":"R. Pollitzer Plague monograph series 22 (World Health Organization Geneva 1954)."},{"key":"e_1_3_1_22_2","unstructured":"For IFA digestive tracts were dissected from individual fleas triturated in PBS containing 0.75% BSA on a microscope slide air dried and fixed in acetone for 20 min. Controls included preparations from uninfected fleas and slides prepared with bacteria grown in brain-heart infusion broth at 21\u00b0C. Slides were incubated with rabbit polyclonal anti-Ymt antiserum in PBS\u20130.75% BSA washed and reincubated with fluorescein-labeled goat anti-rabbit immunoglobulin G."},{"key":"e_1_3_1_23_2","unstructured":"C. R. Eskey V. H. Haas Plague in the Western Part of the United States Public Health Bulletin 254 (U.S. Public Health Service Washington DC 1940) vol. 254."},{"key":"e_1_3_1_24_2","unstructured":"\u00c5. Forsberg P. Cherepanov unpublished data."},{"key":"e_1_3_1_25_2","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1093\/jmedent\/30.1.214","volume":"30","author":"Vaughan J. A.","year":"1993","unstructured":"Vaughan J. A., Azad A. F., J. Med. Entomol. 30, 214 (1993).","journal-title":"J. Med. Entomol."},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/35012500"},{"key":"e_1_3_1_27_2","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.1086\/314456","volume":"178","author":"Hinnebusch B. J.","year":"1998","unstructured":"Hinnebusch B. J., Fischer E. R., Schwan T. G., J. Infect. Dis. 178, 1406 (1998).","journal-title":"J. Infect. Dis."},{"key":"e_1_3_1_28_2","doi-asserted-by":"crossref","first-page":"4310","DOI":"10.1128\/iai.59.12.4310-4317.1991","volume":"59","author":"Donnenberg M. S.","year":"1991","unstructured":"Donnenberg M. S., Kaper J. B., Infect. Immun. 59, 4310 (1991).","journal-title":"Infect. Immun."},{"key":"e_1_3_1_29_2","doi-asserted-by":"crossref","first-page":"5929","DOI":"10.1128\/jb.172.10.5929-5937.1990","volume":"172","author":"Perry R. D.","year":"1990","unstructured":"Perry R. D., Pendrak M. L., Schuetze P., J. Bacteriol. 172, 5929 (1990).","journal-title":"J. Bacteriol."},{"key":"e_1_3_1_30_2","doi-asserted-by":"crossref","first-page":"29497","DOI":"10.1074\/jbc.273.45.29497","volume":"273","author":"Heinrichs D. E.","year":"1998","unstructured":"Heinrichs D. E., Yethon J. A., Amor P. A., Whitfield C., J. Biol. Chem. 273, 29497 (1998).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_31_2","unstructured":"We thank M. Skurnik and C. Whitfield for providing the Y. pseudotuberculosis and E. coli strains; J. M. Musser and F. Gherardini for review of the manuscript; C. Jarrett E. Lorange J. Stuckey and Y. Du for technical assistance; and A. Mora for help with preparing the figures."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.1069972","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T22:35:05Z","timestamp":1704839705000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.1069972"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,4,26]]},"references-count":30,"journal-issue":{"issue":"5568","published-print":{"date-parts":[[2002,4,26]]}},"alternative-id":["10.1126\/science.1069972"],"URL":"https:\/\/doi.org\/10.1126\/science.1069972","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.1006280.78157","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.1006280.83605","asserted-by":"object"}]},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,4,26]]}}}