胸膜肺炎放线杆菌血清8型自转运黏附素基因的克隆测序及功能预测分析

Sequencing and functional analysis of Actinobacillus pleuropneumoniae serotype 8 adhesin gene

为研究胸膜肺炎放线杆菌(APP)三聚体自转运黏附素(TAAs)的功能,以GenBank登录的APP血清5b型自转运黏附素基因5'端的3875bp序列设计引物,通过PCR的方法首次获得APP血清8型运黏附素N端的基因序列片段,测序结果与已知血清型的基因序列和氨基酸推导序列分别进行比对,结果表明与血清7型自转运黏附素N端同源性达到93%,氨基酸推导序列同源性达到97%;与血清5b型自转运黏附素N端同源性达到92%,氨基酸推导序列同源达到100%。经软件分析获得的序列含有与细菌的黏附、聚集和侵入密切相关的Hep_Hag基序,应用马克斯-普朗克研究所的在线分析TAAs的基序和蛋白域的软件daTAA,进行预测并证明所得序列为TAAs,并且具有完整的N段头部序列,有重要功能区具有良好的抗原性。比对的结果为寻找研究APP的定植基序和毒力因素提供了重要基础。

Adhesion is an important pathogenic process for the pathogenesis of bacteria. To study the function of Actinobacillus pleuropneumoniae （APP） autotransporter adhesins （TAAs）, the sequence encoding N part of APP serotype 8 TAAs was amplified by PCR with the primers designed based on the APP serotype 5b transshipment adhesion element gene of 3,875 bp sequence （5＇ end CP000569.1）. The sequence was analyzed by software SMART and PFAM which indicated that the sequence contained Hep_Hag base domain, which was closely related with the the bacterial adhesion, aggregation and intrusive, and the TAAs was predicted in the APP serotype 8 sequence by the Max Planck institute of on-line and adhesion grain protein domain software daTAA analysis. The sequence analysis results provided a important basis for further study of the APP colonization and virulence factors.