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与出血性败血症相关的多杀性巴氏杆菌(B:2)分离株外膜蛋白谱的比较分析。

Comparative analysis of the outer membrane protein profiles of isolates of the Pasteurella multocida (B:2) associated with haemorrhagic septicaemia.

作者信息

Tomer P, Chaturvedi G C, Malik P, Monga D P

机构信息

Department of Veterinary Microbiology, College of Veterinary Sciences, CCS Haryana Agricultural University, Hisar, India.

出版信息

Vet Res Commun. 2002 Oct;26(7):513-22. doi: 10.1023/a:1020212430041.

Abstract

Outer membrane proteins (OMP) of P. multocida (serotype B:2) field isolates (n = 6) and a vaccine strain (P-52) were extracted by a sarkosyl method and characterized using SDS-PAGE and immunoblotting. About 20 polypeptide bands were observed in the profile of the vaccine strain with MW ranging from 16 to 90 kDa and, based on band thickness and intensity of staining, three polypeptides of MW 31, 33 and 37 kDa were considered to be the major OMPs. The profiles of the field isolates showed minor differences when compared with that of the vaccine strain. The OMP of 33 kDa was only expressed by the vaccine strain. Four field isolates expressed an OMP of 39 kDa, which did not appear in the profiles of the remaining two field isolates and the P-52 strain. Similarly, an OMP of 25 kDa was exclusively seen in the profile of a single isolate. By immunoblotting studies, using anti-P. multocida (P-52) whole-cell hyperimmune serum raised in rabbits as well as buffalo immune sera, it became evident that the polypeptide of 37 kDa was the most antigenic OMP in the profiles of all the isolates, including the P-52 strain. Other polypeptides were either weakly antigenic or visible in the profile of only a few of the isolates. The study thus identified the major OMP of P. multocida (B:2) and suggested that this highly antigenic 37 kDa OMP has potential for further protective and immunodiagnostic studies.

摘要

采用十二烷基肌氨酸钠法提取多杀性巴氏杆菌(血清型B:2)的6株野外分离株及1株疫苗株(P-52)的外膜蛋白(OMP),并用SDS-PAGE和免疫印迹法进行鉴定。在疫苗株的蛋白图谱中观察到约20条多肽带,分子量范围为16至90 kDa,根据条带厚度和染色强度,认为分子量为31、33和37 kDa的3种多肽是主要的外膜蛋白。与疫苗株相比,野外分离株的蛋白图谱显示出微小差异。33 kDa的外膜蛋白仅由疫苗株表达。4株野外分离株表达了一种39 kDa的外膜蛋白,其余2株野外分离株及P-52株的蛋白图谱中未出现该蛋白。同样,仅在1株分离株的蛋白图谱中观察到一种25 kDa的外膜蛋白。通过免疫印迹研究,使用兔抗多杀性巴氏杆菌(P-52)全细胞超免疫血清以及水牛免疫血清,结果表明37 kDa的多肽是所有分离株(包括P-52株)蛋白图谱中最具抗原性的外膜蛋白。其他多肽要么抗原性较弱,要么仅在少数分离株的蛋白图谱中可见。该研究鉴定了多杀性巴氏杆菌(B:2)的主要外膜蛋白,并表明这种高度抗原性的37 kDa外膜蛋白具有进一步进行保护性和免疫诊断研究的潜力。

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