Lainson F A, Harkins D C, Wilson C F, Sutherland A D, Murray J E, Donachie W, Baird G D
Moredun Research Institute, Edinburgh, UK.
J Gen Microbiol. 1991 Feb;137(2):219-26. doi: 10.1099/00221287-137-2-219.
Iodination of intact Pasteurella haemolytica serotype A2 cells labelled a sub-set of total cellular proteins. Comparison of the autoradiographic patterns obtained from iodinated cells grown on complete medium and on iron-depleted medium showed that expression of three proteins, of 100, 70 and 35 kDa, respectively, was increased by growth under iron-depleted conditions. Of these proteins, that of 35 kDa had not been reported previously. Like the 100 and 70 kDa proteins, the 35 kDa protein was expressed in natural infections, since it was recognized by antiserum from sheep that had recovered from an experimental infection with P. haemolytica A2. The 35 kDa protein was partially purified by reverse-phase HPLC and was found to be antigenic in both sheep and mice. A monoclonal antibody that was specific for the 35 kDa protein was used to identify the cellular location of the protein by immunoblotting of cell fractions enriched for particular cellular components. This demonstrated that the 35 kDa protein was located mainly in the periplasm.
完整的溶血巴斯德菌A2型细胞的碘化标记了总细胞蛋白的一个亚组。对在完全培养基和缺铁培养基上生长的碘化细胞获得的放射自显影片模式进行比较，结果表明，分别为100、70和35 kDa的三种蛋白质的表达在缺铁条件下生长时增加。在这些蛋白质中，35 kDa的蛋白质以前未曾报道过。与100和70 kDa的蛋白质一样，35 kDa的蛋白质在自然感染中表达，因为它能被从感染溶血巴斯德菌A2的实验感染中恢复的绵羊的抗血清识别。35 kDa的蛋白质通过反相高效液相色谱法进行了部分纯化，并且发现它在绵羊和小鼠中都具有抗原性。一种对35 kDa蛋白质具有特异性的单克隆抗体被用于通过对富含特定细胞成分的细胞组分进行免疫印迹来鉴定该蛋白质的细胞定位。这表明35 kDa的蛋白质主要位于周质中。
