Witchell Timothy D, Coutts Scott A J, Bulach Dieter M, Adler Ben
Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia.
Infect Immun. 2006 Jun;74(6):3271-6. doi: 10.1128/IAI.02000-05.
The enteric, anaerobic spirochete Brachyspira hyodysenteriae is the causative agent of swine dysentery, a severe mucohemorrhagic diarrheal disease of pigs that has economic significance in every major pork-producing country. Recent investigation into potential vaccine candidates has focused on the outer membrane proteins of B. hyodysenteriae. Bhmp39 (formerly Vsp39) is the most abundant surface-exposed outer membrane protein of B. hyodysenteriae; its predicted gene sequence has previously been shown to share sequence similarity to eight genes divided evenly between two paralogous loci. The peptide sequence suggested that Bhmp39 is encoded by one of these genes, bhmp39h. The biological significance of maintaining eight homologous bhmp39 genes is unclear, though it has been proposed that this may play a role in antigenic variation. In this study, real-time, reverse transcription-PCR was used to demonstrate that bhmp39f and bhmp39h were the transcripts most abundantly expressed by B. hyodysenteriae strain B204 cultured under in vitro growth conditions. Mass spectrometry data of the purified 39-kDa membrane protein showed that both Bhmp39f and Bhmp39h were present. Northern blot analysis across predicted Rho-independent terminators demonstrated that the genes of the bhmp39efgh locus result in monocistronic transcripts.
肠道厌氧螺旋体猪痢疾短螺旋体是猪痢疾的病原体,猪痢疾是一种严重的猪黏液出血性腹泻疾病,在每个主要猪肉生产国都具有经济意义。最近对潜在候选疫苗的研究集中在猪痢疾短螺旋体的外膜蛋白上。Bhmp39(以前称为Vsp39)是猪痢疾短螺旋体中最丰富的表面暴露外膜蛋白;其预测的基因序列先前已显示与八个基因具有序列相似性,这些基因平均分布在两个旁系同源位点之间。肽序列表明Bhmp39由这些基因之一bhmp39h编码。维持八个同源bhmp39基因的生物学意义尚不清楚,尽管有人提出这可能在抗原变异中起作用。在本研究中,使用实时逆转录PCR来证明bhmp39f和bhmp39h是猪痢疾短螺旋体B204菌株在体外生长条件下培养时表达最丰富的转录本。纯化的39 kDa膜蛋白的质谱数据表明Bhmp39f和Bhmp39h均存在。对预测的不依赖Rho终止子进行的Northern印迹分析表明,bhmp39efgh位点的基因产生单顺反子转录本。