为了扩增坏死梭杆菌（ Fusobacterium necrophorum ）血凝素相关外膜蛋白基因完整的核苷酸序列和了解该蛋白的分子特征，以坏死梭杆菌AB型菌株的基因组DNA为模板，利用染色体步移技术分别扩增已知序列的5′端旁侧序列和3′端旁侧序列，并测序和拼接，对拼接结果进行生物信息学分析。结果显示，成功获得 F.necrophorum血凝素相关外膜蛋白基因的完整核苷酸序列，全长4247bp ，含有1个3372bp的开放阅读框，编码1123个氨基酸，蛋白质分子质量约为120ku。生物信息学分析表明，该蛋白没有明显的信号肽序列和跨膜结构；该蛋白质第750位～1123位氨基酸残基区域具有较强的抗原性；SMART预测显示，该蛋白存在一个自转运蛋白结构域、血凝活性结构域和多个参与碳水化合物代谢的结构域。上述结果提示，成功获得F.necrophorum血凝素相关外膜蛋白基因的完整核苷酸序列，编码约为120ku的蛋白质，该蛋白质可能是具有血凝活性的外膜自转运蛋白家族的成员，并且该蛋白质的C端可能具有较强的抗原性。%In order to amplify the complete sequence of hemagglutinin -related outer membrane protein gene from Fusobacterium necrophorum and predict the molecular characteristics of the protein .,we used the genomic DNA ,extracted from F .necrophorum ,as the template and ampli-fied it with the methods of 5′end flanking sequence and 3′end flanking sequence genome walking .After connection and sequencing ,the two flanking sequences and the known sequence were spliced together ,consisted of 4247bp .Bioinformatics analysis showed that hemagglutinin -re-lated outer membrane protein gene contained an ORF of 3372bp and encoding 1123 amino acids ,the molecular weight was about 120 ku .The protein had no signal peptide and transmembrane regions and the 750-1123 amino acids residues had the highest antigenicity .In addition ,the protein had a autotransporter domain ,a hemagglutination activity domain and other domains which were involved in carbohydrate metabolism . These results indicated that the complete sequence of hemagglutinin-related outer membrane protein gene from F .necrophorum were success-fully obtained ,the molecular weight was about 120ku .This protein might possess hemagglutination activity and members of autotransporter fami-ly .C-terminal of the protein might be the highest antigenicity .
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