Lockman H A, Curtiss R
Department of Biology, Washington University, St. Louis, Missouri 63130.
Infect Immun. 1990 Jan;58(1):137-43. doi: 10.1128/iai.58.1.137-143.1990.
Nonmotile flagellated (mot) and nonflagellated (fla) mutants of Salmonella typhimurium LT-2 were isolated from a collection of mutants with random Tn10-insertion mutations. Both classes of mutants were resistant to infection by the flagellotropic bacteriophage chi. The nonflagellated (fla::Tn10) mutants did not react with H antigen-specific antisera and did not possess flagella when examined by electron microscopy, and sheared-cell extracts were devoid of flagellin. The nonmotile (mot::Tn10) mutants reacted with H-specific antisera and expressed paralyzed flagella that were indistinguishable from wild-type flagella. The Tn10 insertions in strain LT-2 were mapped to loci in regions II (flh and mot) and III (fli) of the flagellar genes, and the mutations were transduced into the mouse-virulent S. typhimurium strains SR-11 and SL1344. Lack of motility reduced the ability of S. typhimurium to invade Henle cells in vitro, yet the virulence in mice of the nonmotile mutants of SR-11 and SL1344 was unaffected by the inactivity or loss of flagella. Wild-type SR-11 had a 50% lethal dose (LD50) in BALB/c mice following oral (p.o.) challenge of 2.4 x 10(4) CFU. The p.o. LD50 of the SR-11 fli-8007::Tn10 mutant was 4.5 x 10(4) CFU. The mot-8008::Tn10 mutation in SR-11 conferred paralyzed flagella and increased the p.o. LD50 in mice to 2.2 x 10(5) CFU, but this was not statistically significant. A similar increase in the p.o. LD50 was observed when the SL1344 mot-8008::Tn10 mutant was tested in mice. Wild-type SR-11 and the isogenic nonflagellated and nonmotile mutants were equally virulent in mice challenged via intraperitoneal injection.
从一组具有随机Tn10插入突变的突变体中分离出鼠伤寒沙门氏菌LT-2的无运动性鞭毛(mot)和无鞭毛(fla)突变体。这两类突变体均对嗜鞭毛噬菌体chi的感染具有抗性。无鞭毛(fla::Tn10)突变体不与H抗原特异性抗血清发生反应,在电子显微镜检查时也不具有鞭毛,并且剪切细胞提取物中没有鞭毛蛋白。无运动性(mot::Tn10)突变体与H特异性抗血清发生反应,并表达出与野生型鞭毛无法区分的麻痹鞭毛。LT-2菌株中的Tn10插入被定位到鞭毛基因的II区(flh和mot)和III区(fli)的位点,并且这些突变被转导到小鼠致病型鼠伤寒沙门氏菌菌株SR-11和SL1344中。缺乏运动性降低了鼠伤寒沙门氏菌在体外侵袭亨勒细胞的能力,然而SR-11和SL1344的无运动性突变体在小鼠中的毒力不受鞭毛无活性或缺失的影响。野生型SR-11经口服(p.o.)攻击后,在BALB/c小鼠中的半数致死剂量(LD50)为2.4×10⁴CFU。SR-11 fli-8007::Tn10突变体的口服LD50为4.5×10⁴CFU。SR-11中的mot-8008::Tn10突变赋予了麻痹鞭毛,并使小鼠口服LD50增加到2.2×10⁵CFU,但这在统计学上并不显著。当在小鼠中测试SL1344 mot-8008::Tn10突变体时,观察到口服LD50有类似增加。野生型SR-11以及同基因的无鞭毛和无运动性突变体在经腹腔注射攻击的小鼠中具有同等毒力。