摘要
牛种、羊种布鲁氏菌非典型株的菌细胞经机械方法破碎、酶解后用两性离子去污剂提取其外膜蛋白并与光滑型菌株的相应成分进行SDS-PAGE图谱比较,发现两种类型的菌株外膜蛋白图谱有极大的相似之处(即布鲁氏菌光滑型菌株所共有的谱带,非典型株也全具备),但非典型株与光滑型菌株不同的是分子量大于97.4kD的热修饰蛋白电泳谱带发生丢失现象:即这一组的谱带除一条明显变宽外,其余着色变浅或基本消失,当热修饰蛋白在高温SDS系统解聚成单体后,其电泳谱带牛种菌两种类型的菌株基本一致,但对羊种菌非典型株的单体却明显增加了两条谱带,以此显示与光滑型菌株的不同。
Outer membrane proteins were solubilized from 37 smooth strains of different biotypes of B.abortus, B.melitensis and B.suis by sequential extraction of physically disrupted cells with Triton X-100 and a dipolar ionic detergent (Zwittergent-314). Then comparison of the SDS-PAGE profiles among the tested strains was performed. In contrast with previous reports, the reproducible and completed profiles of SDS-PAGE were obtained within three Brucella species. Five principal protein groups with apparent molecular weights of more than 97.4 kd (group 1). near 42.7 kd (group 2), 31-42 kd (group 3), 28-31 kd (group 4) and less than 28 kd (group 5) were observed in every strain. Among them, the group 5 and the group 4 were shared in three species, but gro.up 2 was only shared in the outer membrane protein of B.melitensis and B.suis.
It was revealed that the group 1 protein was actually a heat-modifiable protein. When it was heated in the SDS-system at 100℃, the depolymerization of group 1 protein occured, turned into its monomer (group 3), and its molecular weight was about 1/3 of the group 1 protein. It's important that the significant differenciation of the pattern of SDS-PAGE profiles of the monomer among the three tested species was confirmed. One band of the monomer (near 31 kd) was appeared in B.melitensls; two bands, closed-space and with smaller dispersion, were representative monomer of B.abortus; and another two broad bands, which migrate rather slowly, were the monomer of B.suis.
In brief, to confirm Brucella species by using the pattern of SDS-PAGE profiles, firstly the bands near 42.7 kd region (group 2) disappeared obviously at normal temperature in SDS system, the tested strain must belong to B.abortus species. If these bands were observed significantly in same conditions, the tested strain must be B.melitensis or B.suis species. Besides, the special pattern of the monomer of SDS-PAGE profiles in representing each of species could further establesh the species of the tested strains.
出处
《地方病通报》
1991年第1期1-4,131-132,共4页
Endemic Diseases Bulletin