Hackett J, Wyk P, Reeves P, Mathan V
J Infect Dis. 1987 Mar;155(3):540-9. doi: 10.1093/infdis/155.3.540.
A cosmid bank of the DNA (including cryptic plasmid DNA) of a virulent strain of Salmonella typhimurium was prepared in Escherichia coli K12, and clones that contained cryptic plasmid DNA were detected by probing. Two such clones expressed a new outer membrane protein of 11 kilodaltons (kDa) and were serum resistant (E. coli K12 is serum sensitive). The gene encoding the 11-kDa protein was subcloned in a 2.1-kilobase fragment and shown to mediate serum resistance in both E. coli K12 and a cryptic plasmid-free (serum-sensitive) strain of S. typhimurium. The cryptic plasmid-free S. typhimurium strain did not express normal lipopolysaccharide, but introduction of the 11-kDa protein gene into the strain rendered the strain serum resistant without restoration of normal lipopolysaccharide synthesis. The 11-kDa protein gene was not sufficient to restore either macrophage resistance or virulence to a cryptic plasmid-free strain of S. typhimurium.
构建了鼠伤寒沙门氏菌强毒株DNA(包括隐蔽质粒DNA)的黏粒文库,并将其导入大肠杆菌K12中,通过探针检测含有隐蔽质粒DNA的克隆。两个这样的克隆表达了一种新的11千道尔顿(kDa)的外膜蛋白,并且具有血清抗性(大肠杆菌K12对血清敏感)。编码11-kDa蛋白的基因被亚克隆到一个2.1千碱基的片段中,并显示在大肠杆菌K12和无隐蔽质粒(血清敏感)的鼠伤寒沙门氏菌菌株中均介导血清抗性。无隐蔽质粒的鼠伤寒沙门氏菌菌株不表达正常的脂多糖,但将11-kDa蛋白基因导入该菌株可使其具有血清抗性,而不会恢复正常的脂多糖合成。11-kDa蛋白基因不足以恢复无隐蔽质粒的鼠伤寒沙门氏菌菌株对巨噬细胞的抗性或毒力。