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部分菌株中与毒力相关的免疫原性蛋白的测定

Determination of virulence associated immunogenic proteins in some of strains.

作者信息

Yesiltas Meryem Cansu, Altinok Ilhan, Ozturk Rafet Cagri

机构信息

Department of Fisheries Technology Engineering, Surmene Faculty of Marine Sciences, Karadeniz Technical University, Trabzon, Turkey.

出版信息

Vet Res Forum. 2019 Spring;10(2):101-107. doi: 10.30466/vrf.2018.89527.2164. Epub 2019 Jun 15.

Abstract

Lactococcosis disease incident caused by has been increased with increasing aquaculture productions and outbreaks of the disease have become a threat on farmed species. To prevent lactococcosis, inactivated vaccine has been used, however, it only provides protection when given by injection. Other than inactivated vaccine, various vaccines such as subunit vaccines can be developed. In the present study, total protein profile of 43 strains of isolated from fish, milk and cheese by SDS-PAGE and virulence associated immunogenic proteins of strains using western blot with hyper-immune rabbit sera were determined. After analyzing whole-cell lysate protein of strains with SDS-PAGE, protein bands were ranged between 8.00 and 140.00 kDA. Among strains, variable protein bands were ranged between 17.00 and 48.00 kDa with some variability in the staining intensity of the protein bands and formed in 6 clusters. The immunogenic protein bands were ranged between 25.00 - 75.00 kDa. Only a variable and highly immunogenic protein band was observed between 40.00 and 45.00 kDa. Most of the strain including Lgper had 44.00 kDa immunogenic protein while nonvirulent ATCC strain had 42.50 kDA immunogenic protein. Predominant immuno-reactive proteins encoded by genes can be used as a subunit vaccine.

摘要

由[未提及的病原体]引起的乳球菌病事件随着水产养殖产量的增加而增多,该病的爆发已对养殖物种构成威胁。为预防乳球菌病,已使用灭活疫苗,然而,它仅在注射时提供保护。除了灭活疫苗外,还可以开发各种疫苗,如亚单位疫苗。在本研究中,通过SDS-PAGE测定了从鱼、牛奶和奶酪中分离出的43株[未提及的菌株]的总蛋白谱,并使用超免疫兔血清通过western blot测定了菌株的毒力相关免疫原性蛋白。用SDS-PAGE分析菌株的全细胞裂解物蛋白后,蛋白条带范围在8.00至140.00 kDa之间。在菌株中,可变蛋白条带范围在17.00至48.00 kDa之间,蛋白条带的染色强度存在一些差异,并形成6个簇。免疫原性蛋白条带范围在25.00 - 75.00 kDa之间。仅在40.00至45.00 kDa之间观察到一条可变且高度免疫原性的蛋白条带。包括Lgper在内的大多数菌株具有44.00 kDa的免疫原性蛋白,而无毒力的ATCC菌株具有42.50 kDa的免疫原性蛋白。由基因编码的主要免疫反应性蛋白可作为亚单位疫苗。

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