迟钝爱德华氏菌外膜蛋白基因工程表达产物的免疫原性
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福建省自然科学基金项目(2015J01143); 福建省教育厅项目(JA15283)


Immunogenicity of outer membrane protein expressed by gene engineering of Edwardsiella tarda
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    摘要:

    构建迟钝爱德华氏菌外膜蛋白重组表达载体后,表达纯化获得分子质量约53 ku的外膜蛋白。将100尾日本鳗鲡均分为2组,分别腹腔注射牛血清白蛋白(BSA,0.5 mg/mL)和迟钝爱德华氏菌外膜蛋白(Omp,0.5 mg/mL)0.2 mL/尾。于免疫后第7、14和28天采集鳗鲡血液、粘液、肝脏和肾脏,测定各时间点的补体活性、溶菌酶活性、全血细胞转化水平和血清抗体效价。结果发现,外膜蛋白组鳗鲡的补体活性于免疫后第7天极显著高于对照组;其抗体效价于第14天和第28天显著或极显著高于对照组。免疫后第28天以迟钝爱德华氏菌、嗜水气单胞菌和创伤弧菌(1.0×107 cfu)腹腔注射感染鳗鲡。结果发现,外膜蛋白组鳗鲡对迟钝爱德华氏菌、嗜水气单胞菌和创伤弧菌的相对免疫保护率分别为71.4%、50.0%和12.5%。表明鳗鲡注射迟钝爱德华氏菌基因工程表达外膜后显著提高了其对迟钝爱德华氏菌感染的抵抗力,对嗜水气单胞菌的感染也有明显的交叉保护效果,但对创伤弧菌的交叉保护效果较弱。

    Abstract:

    The expressed outer membrane protein (Omp,about 52 ku) of Edwardsiella tarda was purified after the recombinant expression vector was constructed,transferred into E.coli BL21 and induced.A total of 100 healthy Japanese eels (Anguilla japonica) was divided into 2 groups and intraperitoneally injected (i.p) with bovine serum albumin (BSA,0.5 mg/mL) and the outer membrane protein (Omp,0.5 mg/mL),respectively.On 7,14 and 28 d post-vaccination,the blood,skin mucus,liver and kidney of eels were collected to evaluate the lysozyme and complement activities,stimulation index and specific antibody titers.The complement activities in the Omp group were extremely significantly(P<0.01) higher than that of the BSA group on 7 d.The specific antibody titers in the Omp group were significantly (P<0.05 or P<0.01) higher than that of the BSA group on 14 d and 28 d.The lysozyme activities and stimulation indexes showed no significant difference (P>0.05) between two groups.The relative percent survival (RPS) of Omp were 71.4%,50.0% and 12.5% respectively after challenged by E. tarda,A. hydrophila or V. vulnificus.These results suggested that the recombinant expressed Omp could significantly improve the immune function and resistance of A. japonica against E. tarda.It also had significant cross protection against A. hydrophila,but had weak cross protection against V. vulnificus.

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段丽华,冯建军,郭松林,陈义航,阮彩章.迟钝爱德华氏菌外膜蛋白基因工程表达产物的免疫原性[J].华中农业大学学报,2016,35(5):97-104

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  • 收稿日期:2015-11-10
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  • 在线发布日期: 2016-07-27
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