Preparing the ISCOMs of Aeromonas hydrophila β-hemolysin expressed in E.coli
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摘要: 嗜水气单胞菌β-hemA 重组菌 PCB 的培养上清液经硫酸铵梯度沉淀粗提毒素蛋白质,并在pH2.5的柠檬酸缓冲液里酸化处理以暴露其疏水区,经Mega-1O裂解后与 Quil A 结合制备 ISCOMs,电镜下可观察到30～40nm 的多面体的笼格状颗粒结构。经鳗鱼的免疫保护试验证实,ISCOMs组(每尾40~80pg)保护率高达100%,同剂量的溶血素+弗氏佐剂组可达62.5%,未加佐剂的溶血素只有50%。
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关键词:
- 嗜水气单胞菌 /
- 免疫刺激复合物(ISCOMs) /
- 免疫
Abstract: A method for preparing the immuno-stimulating complexes(ISCOMs)with theβ-hemolysin(β-HemA) and Quil A was described,in which the β-hemolysin of Aeromonas hydrophila was extracted by ammonium sulphate gradient precipitation and pretreated with an acidic pH(2.5)to reveal hydrophobic regions,and then was mixed with the Quil A and other reagents.The ISCOMs was a cagelike Structure with a diameter of 30—40 nm by negative contrast electron microscopy.Several groups of eel were immunized with β-HemA-ISCOMs,β-HemA-CFA and same amount of β-hemolysin without adjuvant,respectively,and followed by challenge with a virlent bacterial strain TPS-30.The protecion rates of that were 100%,62.5%,and 50%,respectively.The result indicated that the eel immunized with β-HemA-ISCOMs were effectively protected against the challenge.-
Key words:
- Aeromonas hydrophila /
- Immune stimulating complexes (ISCOMs) /
- Immunology
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[1] 陈月英,钱冬,沈智华,等.淡水鱼类细菌性败血症菌苗浸浴免疫研究[J]海洋与湖沼,1998,29(6):597-603. [2] Akerblom L,Stromstedt K,Hoglund S,et al.Formation and characterization of FeLV ISCOMs[J].Vaccine,1989,7(2):142-146. [3] 龚晖,林天龙,杨金先,等.欧鳗源嗜水气单胞菌β-溶血素基因的序列分析[J].福建农业学报,2003,18(1):29-33. -
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