摘要
目的研究钩端螺旋体外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎ，ＯＭＰ）的结构和抗原特征，对不同种属钩端螺旋体疏水外膜蛋白进行了比较研究。方法用ＴｒｉｔｏｎＸ－１１４（ＴＸ－１１４）抽提１０株不同种属钩体疏水外膜蛋白，用不连续缓冲系统制备５％浓缩胶和１２％的分离胶，行ＳＤＳ－ＰＡＧＥ电泳，在恒压下１８０Ｖ电泳５小时，考马斯亮兰Ｇ－２５０染色。用抗０１７外膜血清、０１７ＭｃＡｂ行免疫印迹分析，对其疏水外膜蛋白的免疫特征进行比较和分析。结果不同种属钩体疏水ＯＭＰ，均出现４－７条主要蛋白区带，并存在较大差别。致病性问号状钩体与非致病的双曲钩体及细丝体几乎无相同的蛋白主带，而分子量为３９０００的蛋白主带为问号状钩体各株共有，且各株此区带与问号状钩体０１７株特异性抗体出现强弱不等的免疫反应；不同群型０１７株和ＨｏｎｄＵｔｒｅｃｈｔⅣ株带型相似程度较大，而同型不同株０１７株（高毒）和６０１株（低毒）两株有相似主带，也有各自不同的主带。结论不同种属钩体疏水ＯＭＰ结构有较大差异，同种、同群、同型间相似性较大，但也有明显变异。分子量为３９０００疏水ＯＭＰ（ＯｍｐＬ３９）是问号状钩体共有的蛋白抗原，在筛选钩体毒力基因、保?
Objective To study the differences of OMPs of Leptospira of different genera in reference to their antigenicities and molecular characteristics. Methods\ The outer membrane proteins of the Leptospira of different species have been extracted with TX 114.SDS PAGE was used for analyses of different OMPs.5% concentrated gel and 12% separating gel were prepared with discontinued buffer systems.The electrophoresis were done at 180V for 5 hrs,and the gels were stained with Coomassie brilliant blue G 250. Results \ The structures of the hydrophobic OMPs of L.interrogans,L.biflexa and Leptonema illini were apparently different,all the OMPs of 10 strains had 4 7 major protein bands but the profile patterns were not the same.All strains of L.interrogans had the protein of 39 kD and 6 of 8 strains of L.interrogans gave positive results in immunoblotting using strain 017 antibody, while hydrophobic OMPs of L.biflexa and Leptonema illini were negative,and some major bands were same in strains 017(Serogroup Icterohaemorrhagiae) and Hond Utrecht IV(Serogroup Canicola)of L.interrogans ,both L.interrogans serovar Lai strain 017(high virulent) and Lai strain(low virulent) contained similar major bands and some bands were unique to strain 017 or Lai. Conclusions It suggests that changes of structural gene of hydrophobic OMPs might result in changes of virulence,pathogenicity and immunogenicity of different genera and species of Leptospira. The hydrophobic OMP of 39kD was one of the common protein antigen in L.interrogans and might be important for screening virulent genes,immunoprotective antigens and identification of Leptospria.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1998年第3期189-193,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金
卫生部留学回国人员科研启动基金
关键词
钩端螺旋体
膜蛋白
抗原性
Leptospira Hydrophobic Membane protein\ \ Antigen
