Mills S D, Bradbury W C
Infect Immun. 1984 Feb;43(2):739-43. doi: 10.1128/iai.43.2.739-743.1984.
Two techniques were used to isolate outer membrane proteins from Campylobacter jejuni, EDTA-lysozyme extraction and sodium-N-lauroylsarcosinate (Sarkosyl) solubilization. The protein profiles of the two preparations were similar, with a few additional bands in the EDTA-lysozyme preparations. The major outer membrane protein was 43,000 (43K) daltons, and there were 8 to 10 minor bands ranging from 92K to 14K daltons. There was no difference in the protein profile of a strain causing an infection (strain 17) and the resulting stool isolate (strain 17J). Sera collected before the infection and during the acute and convalescent stages were used with Western blotting and immunoautoradiographic techniques to determine the antigenicity of outer membrane proteins. A number of antigenic proteins were detected before the infection by their reaction with preinfection serum (61K, 51K, 43K, 40K, 34K, and 31K daltons), and three additional bands appeared during the infection when acute and convalescent sera were used (92K, 56K, and 19K daltons). Furthermore, an area of the gel at less than 14.4K daltons that did not stain with Coomassie brilliant blue became visible in the immune blots when the convalescent serum was used.
采用两种技术从空肠弯曲菌中分离外膜蛋白,即乙二胺四乙酸 - 溶菌酶提取法和N - 月桂酰肌氨酸钠(十二烷基肌氨酸钠)增溶法。两种制备物的蛋白质谱相似,不过在乙二胺四乙酸 - 溶菌酶制备物中有几条额外的条带。主要的外膜蛋白为43,000(43K)道尔顿,还有8至10条次要条带,分子量在92K至14K道尔顿之间。引起感染的菌株(菌株17)和由此产生的粪便分离株(菌株17J)的蛋白质谱没有差异。在感染前、急性期和恢复期收集的血清用于蛋白质印迹法和免疫放射自显影技术,以确定外膜蛋白的抗原性。在感染前通过与感染前血清的反应检测到一些抗原性蛋白(61K、51K、43K、40K、34K和31K道尔顿),当使用急性期和恢复期血清时,在感染期间又出现了另外三条条带(92K、56K和19K道尔顿)。此外,当使用恢复期血清时,在免疫印迹中,凝胶上小于14.4K道尔顿且未被考马斯亮蓝染色的区域变得可见。