首页|维氏气单胞菌外膜蛋白OmpH功能预测及免疫原性分析

维氏气单胞菌外膜蛋白OmpH功能预测及免疫原性分析

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对维氏气单胞菌OmpH基因编码蛋白(OmpH)进行生物学特性分析,预测其B细胞和T细胞优势抗原表位,并对该基因进行克隆和原核表达,分析其表达产物的免疫原性.结果表明:维氏气单胞菌OmpH基因全长513 bp,编码的蛋白有跨膜区,为亲水性外膜蛋白,二级结构主要为α-螺旋;抗原表位预测结果表明,OmpH蛋白的B细胞潜在优势抗原肽段为112~114位氨基酸,含有7个辅助性T细胞(Th)表位及6个细胞毒性T细胞(CTL)表位;构建Escherichia coli pColdTF-ompH原核表达菌株,成功表达OmpH蛋白,最佳诱导时间为24 h,最佳诱导剂终浓度为1.0 mmol/L;Western blot分析结果表明,重组菌诱导后表达的OmpH蛋白可以被兔抗OmpH蛋白血清识别,即OmpH蛋白具有良好的免疫原性.
Function Prediction and Immunogenicity Analysis of Outer Membrance Protein OmpH of Aeromonas versonii
The biological characteristics of the protein(OmpH)encoded by OmpH gene of Aeromonas veronii were analyzed,the dominant epitopes of B and T cells were predicted,the gene was cloned and prokaryotic expressed,and the immunogenicity of the expression product was analyzed in the study.The results showed that ompH gene of Aeromonas veronii was 513 bp in length,and the encoded protein had a transmembrane region,which was a hydrophilic outer membrane protein,and its secondary structure was mainly α-helix.Antigenic epitope prediction results showed that the B cell potential dominant antigenic peptide fragment of OmpH protein was 112-114 amino acids,and 7 Th and 6 CTL cell epitopes were predicted.Escherichia coli pColdTF-ompH prokaryotic expression strain was constructed,and OmpH protein was successfully expressed.The optimal induction time was 24 h,and the final concentration of the best inducer was 1.0 mmol/L.The results of Western-blot analysis showed that OmpH protein expressed after induction by recombinant bacteria could be recognized by rabbit anti-OmpH protein serum,which indicated that OmpH protein had good immunogenicity.

Aeromonas veroniiouter membrane proteinOmpH geneprokaryotic expressionim-munogenicity

李洪锦、赵桐、曹立男、康元环

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吉林农业大学动物医学院/动物科学技术学院,长春 130118

动物生产及产品质量安全教育部重点实验室,长春 130118

山东大学海洋学院,威海 264209

维氏气单胞菌 外膜蛋白 OmpH基因 原核表达 免疫原性

吉林省自然科学基金项目

20210101050JC

2024

经济动物学报
吉林农业大学

经济动物学报

CSTPCD
影响因子:0.822
ISSN:1007-7448
年,卷(期):2024.28(2)
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