排序方式: 共有35条查询结果,搜索用时 15 毫秒
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为分离纯化奶牛腐蹄病坏死杆菌,分析其与其他菌株的亲缘关系,本研究利用坏死杆菌白细胞毒素特异性引物,对奶牛腐蹄病病牛蹄部拭子样品进行了PCR检测,利用厌氧培养基对PCR检测阳性样品进行了坏死杆菌的分离培养,以分离的坏死杆菌基因组DNA为模板,对白细胞毒素基因进行了克隆和序列分析。结果显示,9份奶牛腐蹄病病牛蹄部拭子样品PCR检测结果均为阳性,对其中一份样品中的坏死杆菌进行分离培养,获得了纯培养物,命名为bFR13-1。坏死杆菌bFR13-1菌株白细胞毒素基因测序结果显示,与GenBank已发表的H05、A25和B35菌株的白细胞毒素基因在核苷酸水平的同源性分别为98.40%、98.35%和90.79%,推导氨基酸的同源性分别为97.7%、97.6%和89.0%。进化树分析结果显示,坏死杆菌bFR13-1菌株白细胞毒素与H05菌株的同源性最高,bFR13-1菌株与H05菌株和A25菌株呈较近亲缘关系。结果表明,不同坏死杆菌分离株的白细胞毒素呈现一定的变异性,这种变化是否与坏死杆菌致病性相关,值得深入研究。 相似文献
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Septic osteomyelitis of the temporomandibular joint (TMJ) is rarely seen in equine medicine with few reported cases, and none reported involving Fusobacterium necrophorum. This report describes the diagnosis and successful treatment of septic osteomyelitis of the temporomandibular joint (TMJ) in an 11-year-old Australian Stockhorse mare in Queensland, Australia. 相似文献
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S. Saginala T. G. Nagaraja Z. L. Tan K. F. Lechtenberg M. M. Chengappa P. M. Hine 《Veterinary research communications》1996,20(6):493-504
The serum antileukotoxin antibody response and protection against subsequent experimental challenge with Fusobacterium necrophorum were investigated in 30 steers vaccinated with crude F. necrophorum leukotoxoid. Culture supernatant of F. necrophorum, strain 25, containing leukotoxoid was concentrated. The steers were assigned randomly to six groups (n=5): PBS control with Stimulon adjuvant; vaccinated with concentrated supernatant diluted to provide 2.5, 5.0, 10.0, or 20.0 ml with the watersoluble Stimulon adjuvant; and 5.0 ml with the Ribi oil-emulsion adjuvant. The steers were injected subcutaneously on days 0 and 21. Blood samples were collected at weekly intervals to monitor serum antileukotoxin antibody titres. On day 42, all the steers were challenged intraportally with F. necrophorum culture. Three weeks later (day 63), the steers were killed and necropsied for examination of their livers and assessment of protection. Steers vaccinated with crude leukotoxoid tended to have higher antileukotoxin titres than the controls, but the difference was not significant. Also, the antibody titre did not appear to be dose-dependent. In the control group, 3 out of 5 steers developed liver abscesses. The incidence of liver abscesses in steers vaccinated with Stimulon adjuvant was not dose related; however, only 8 of the 25 vaccinated steers developed abscesses. None of the steers vaccinated with the 5.0 ml dose with Ribi had any abscesses. Evidence for a relationship between antileukotoxin antibody and protection was shown by the lower titre in those steers that developed abscesses compared to those that did not. It was concluded that antileukotoxin antibody titres probably provided some degree of protection against experimentally induced liver abscesses, but further dose-titration studies using Ribi or possibly another more effective adjuvant will be needed to confirm this.Abbreviations BHI
brain-heart infusion
- CFU
colony-forming units
- ELISA
enzyme-linked immunosorbent assay
- MTT
3-[4,5-(dimethylthiazol-2-yl)]-2,5-diphenyltetrazodium] bromide
- PBS
phosphate-buffered saline
- PMN
polymorphonuclear neutrophils 相似文献
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坏死梭杆菌是动物和人的各种坏死化脓感染的条件性致病菌.坏死梭杆菌的白细胞毒素是一种高度不稳定性分泌蛋白,被认为是主要的毒力因子.坏死梭杆菌白细胞毒素基因的开放阅读框(lktAORF)包括9 726 bp,编码3 241个氨基酸,总分子质量为336 ku的蛋白,且与其他细菌的细胞毒素没有任何相似的序列.覆盖在整个坏死梭杆菌lktA ORF上的5个短的重叠的多肽分别是BSBSE,SX,GAS,SH和FINAL,将它们在大肠埃希菌中表达,所有的多肽都有免疫原性,但GAS引起最小的抗体反应,BSBSE和SH对坏死梭杆菌攻击诱导产生了很强的保护力,比坏死梭杆菌的培养上清内全长活性lkt或无活性上清的保护性要好得多. 相似文献
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WU Hong-chao LIU Xiao-ying FENG Er-kai ZHU Yan-zhu WAN Hong-li FENG Zhuo YAN Xi-jun CHEN Li-zhi 《中国畜牧兽医》2015,42(11):2843-2849
In order to analyze the immunogenicity of 120 ku hemagglutinin-related outer membrane protein from F.necrophorum,specific primers were designed to amplify the 3 truncated overlapping gene fragments covering the whole open reading frame (ORF) based on the result of antigenic epitope analysis.The gene fragments were ligated into the prokaryotic expression vector to construct pET-28a-p1,pET-32a-p2 and pET-32a-p3,respectively.Then recombinant plasmid was induced expression in E.coli.The SDS-PAGE results showed that the expression of recombinant proteins P1,P2 and P3,molecular mass of the expressed proteins were about 48,59 and 62 ku,respectively.Western blotting indicated that the recombinant proteins could be recognized by antiserum against F.necrophorum from mouse.Humoral immunity response against recombinant proteins was detected in the immunized rabbits.Altogether,these findings suggested that 120 ku hemagglutinin-related outer membrane protein from F.necrophorum had good immunogenicity,and this study provided a reference for further research on genetic engineering subunit vaccine of F.necrophorum. 相似文献
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GAO Jing LI Chun-qiu YAO Shuang ZHANG Li-chun WANG Xin-yu WANG Zhi-hui ZHAO Xi-wen WEI Shan WANG En-yu SUN Dong-bo WU Rui GUO Dong-hua 《中国畜牧兽医》2015,42(9):2308-2312
For separation and purification of Fusobacterium necrophorum of cow footrot, and analysis of genetic relationship with other strains, the hoof ministry swab samples were detected by PCR based on specific primers of leukotoxin gene, and genomic DNA were isolated from PCR positive samples of Fusobacterium necrophorum culturing in anaerobic medium.The genes of leukotoxin were cloned and sequenced.The results showed that nine of hoof ministry swab samples were all PCR positive samples, and we obtained Fusobacterium necrophorum pure culture from one of the samples which named bFR13-1.The gene sequencing results indicated that the homologies of leukotoxin gene nucleotide sequence of bFR13-1 strain compared with H05, A25 and B35 strains from GenBank were 98.40%, 98.35% and 90.79%, respectively, and the homologies of deduced amino acid sequence were 97.7%, 97.6% and 89.0%, respectively.Phylogenetic tree analysis results showed that leukotoxin gene of Fusobacterium necrophorum bFR13-1 and H05 had high homology and bFR13-1, H05 and A25 showed a close genetic relationship.The result indicated that leukotoxin showed variability between different Fusobacterium necrophorum isolated strains, and it was worth to study whether this change and pathogenicity of Fusobacterium necrophorum were related. 相似文献
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牛腐蹄病坏死梭杆菌H05菌株白细胞毒素基因的原核表达及其免疫活性分析 总被引:1,自引:1,他引:1
参考羊腐蹄病坏死梭杆菌白细胞毒素蛋白的抗原表位基因序列,利用DNAStar软件预测了牛腐蹄病坏死梭杆菌白细胞毒素蛋白的5个抗原表位区,设计5对在上游和下游含有特异性限制性内切酶的引物,以牛腐蹄病坏死梭杆菌H05菌株白细胞毒素基因阳性质粒pMD18-T-lkrA为模板,PCR扩增了预测的5个抗原表位区基因,分别命名为PL1、PL2、PL3、PL4和PL5,将其定向克隆到原核表达载体pGEX-6p-1和pPROEX HTa后转化E.coli BL21(DE3),37℃条件下,用IPTG诱导表达,结果PL1、PL2、PL4和PL5在pGEX-6p-1中获得了表达,而PL3在pPROEX HTa中获得了表达。Westem blot试验结果表明,牛腐蹄病坏死梭杆菌H05菌株白细胞毒素蛋白5个抗原表位区的重组蛋白PL1、PL2、PL3、PL4和PL5均与坏死梭杆菌多克隆血清反应。 相似文献