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用RT-PCR方法从水泡性口炎病毒(vsv)扩增印第安纳(Indiana)和新泽西(New Jersey)血清型核蛋白N基因,分别克隆至pMD20-T载体进行序列鉴定及生物信息学分析.构建重组表达质粒VSV-IN-pBCX和VSV-NJ-pB-CX,并经SDS-PAGE和Western-blot鉴定表明2种血清型病毒核衣壳蛋白N基因在BL21(DE3)宿主菌中成功表达,重组蛋白相对分子质量约为82 000,并能够与各自对应血清型阳性血清反应.这表明2个重组蛋白具有良好的抗原性,可作为用于建立水泡性口炎血清学诊断方法的诊断抗原. 相似文献
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重组水泡性口炎病毒基质蛋白制备及ELISA检测方法的建立 总被引:1,自引:0,他引:1
水泡性口炎由水泡性口炎病毒(VSV)引起。VSV基因组编码5个结构蛋白,其中基质蛋白(matrix protein,M)是其重要毒力因子,可阻遏宿主细胞mRNA由胞核向细胞质的转运,并抑制感染病毒的细胞产生I型干扰素IFNα/β,使病毒得以快速繁殖。本实验克隆了印第安那型VSV的M基因,在大肠杆菌表达系统中表达制备重组M蛋白,以M蛋白为抗原建立了检测特异M蛋白抗体的间接ELISA方法。采用建立的ELISA方法分析了VSV感染小鼠体内M蛋白抗体的变化规律。 相似文献
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动物水泡性口炎病毒的抗原性 总被引:1,自引:0,他引:1
动物水泡性口炎病毒(VSV)引起动物水泡性疾病.VSV有两个主要抗原群是核酸蛋白和G蛋白抗原.G蛋白因为能产生中和性抗体被认为是在发动抑制病毒感染方面起主要作用.已经证实VSV 的New Jersey 株中产生独立的中和性单克隆抗体的抗原决定蔟分布在G 蛋白的193~289氨基酸之间. 相似文献
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Romanutti C Bruttomesso AC Castilla V Bisceglia JA Galagovsky LR Wachsman MB 《Veterinary journal (London, England : 1997)》2009,182(2):327-335
In this work the antiviral activity of 20 dehydroepiandrosterone (DHEA) analogs with different substituents at positions C-3, C-15, C-16 and C-17 were evaluated against vesicular stomatitis virus (VSV) in Vero cell cultures. The selectivity indexes (SI) obtained with DHEA and epiandrosterone (EA) were 50 and 72.6, respectively. The work showed that the compounds 21-norpregna-5,17(20)-dien-3β,16α-diyl-diacetate, 17,17-ethylendioxyandrostan-5,15-dien-3β-ol and 3β-hydroxypregn-17(20)-en-16-one had higher SI values than ribavirin, which was used as a reference drug. The antiviral mode of action of DHEA was also investigated against VSV replication in Vero cells, and time of addition experiments showed that DHEA mainly affected a late event in the virus growth cycle. Analysis of RNA and protein synthesis indicated that DHEA adversely affected positive strand RNA synthesis and viral mature particle formation. 相似文献
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This study was aimed to clone and express the specific antigen nucleoprotein (N) of vesicular stomatitis virus (VSV), and then purify and analyze its immunogenicity. Based on published VSV genome N gene sequence in GenBank, two kinds of N genes of VSV with different serotypes were synthesized, respectively. After sequence analysis, one pair of specific primers was designed and synthesized, N gene fragment with about 1 300 bp length was amplified by PCR, and subcloned into pCold Ⅰ expression vector. The recombinant N protein was induced with IPTG and purified by Ni-NTA. The results of SDS-PAGE showed that the N gene was successfully expressed in E. coli and the molecular weight of protein was 50 ku; The results of Western blotting showed that this recombined protein specifically reacted with polyclonal antibody serum of VSV. The recombinant vector with VSV-IND and VSV-NJ were successfully constructed, and the N protein was solubly expressed in E. coli, and the purified protein demonstrated promising immunogenicity. 相似文献
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Matrix蛋白和G蛋白双位点突变型水泡性口炎病毒的构建及生物学活性观察 总被引:1,自引:0,他引:1
重组水泡性口炎病毒(Vesiclar stomatitis virs,VSV)是一种具有良好应用前景的病毒载体,可应用于制备疫苗和治疗肿瘤,但该载体仍存在安全性问题,高剂量注射实验动物可导致产生神经毒性.为减低乃至去除野生型VSV的致病性,作者针对VSV毒力因子,构建了Matrix蛋白和G蛋白双位点突变型重组VSV,该病毒敲除了原来Matrix蛋白的第51位精氨酸和G蛋白C末端的28个氨基酸.相比野生型VSV,新病毒的致病性显著降低.试验也证实这个致弱的病毒是由于2种不同弱化机理共同作用所致,即Ⅰ型干扰素作用和复制能力减低.新型VSV病毒载体有希望成为一个更安全、有效的疫苗载体. 相似文献
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Jinling Wang Yu Wang Yulin Ding Jirintai Suljid Wenlong Wang 《Journal of veterinary diagnostic investigation》2021,33(2):345
A 1-mo-old reticulated giraffe had progressive anorexia and died at the Ordos Zoo. Autopsy revealed necrotic stomatitis with severe bilateral necroulcerative lesions at the base of the tongue and of the cheeks near the commissures of the mouth. There was also severe bilateral confluent bronchopneumonia with a pronounced bronchial pattern and multifocal fibrinous pleuritis. Histologically, there was serofibrinous-suppurative bronchopneumonia with necrosuppurative bronchiolitis and necrotic arteritis. Filamentous bacteria with morphology consistent with Fusobacterium necrophorum were observed at the advancing edge of the necrotic tissue in the tongue and cheeks, as well as in the affected alveolar spaces and bronchioles. Aggregates of slender, gram-negative, rod-like or filamentous bacteria were identified in the lung impression smear. PCR results of 16S rDNA of the tongue and lung lesions had 100% homology to the F. necrophorum subsp. funduliforme B35 sequence (). The gross, histologic, Gram stain, and PCR product sequencing features in our case were consistent with oral and pulmonary necrobacillosis in ruminants, a rare disease of giraffes. EF447425.1相似文献
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P. J. Johnson A. M. LaCarrubba N. T. Messer S. E. Turnquist 《Equine Veterinary Education》2012,24(4):182-186
Two horses were presented for diagnosis of lethargy, salivation, halitosis, dysphagia, bleeding ulcerative glossitis, gingivitis, and focal bleeding ulceration along the mucocutaneous junction of the labial commissures. In both instances, ulceration was attributed to foxtail awns (Setaria geniculata) based on microscopic examination of representative tissue biopsies and identification of numerous foxtail seed heads in representative samples of the available hay. The horses recovered and the lesions resolved completely following removal of the affected source of hay. 相似文献