共查询到19条相似文献,搜索用时 233 毫秒
1.
2.
3.
Mx(myxovirus resistance)蛋白是干扰素诱导产生的具有GTP酶活性的广谱抗病毒蛋白,可抑制病毒复制。本文综述了Mx蛋白结构及其功能,阐述了其抗病毒的作用机制,并对Mx蛋白在抗病毒免疫蛋白方面的应用前景进行了探讨。 相似文献
4.
5.
6.
随着我国水禽业的飞速发展,其集约化程度的提高、饲养条件的恶化或改变、抗生素的药物残留问题、病毒抗原漂移、超强毒株或新型传染病的出现及母源抗体的干扰等系列问题,在生产上引起大量的死亡或造成亚健康状态,进而造成巨大经济损失。这迫使我们要探索新的防疫途径和重新重视抗病育种的研究。Mx蛋白是干扰素(IFN)在体内主要诱导合成的一种特异的抗病毒蛋白,如小鼠Mx1蛋白证明能抑制流感病毒、多里病毒、索戈托病毒的复制;小鼠Mx2蛋白能抑制棒状疱疹性口炎病毒的复制;人MxA蛋白能抑制麻疹副粘病毒、棒状疱疹性口炎病毒、布尼亚病毒、白蛉病毒、汉坦病毒、3型腺病毒的复制”。本文介绍了国内外Mx蛋白的发现历程、结构、抗病毒活性并探讨了Mx蛋白在水禽抗病育种和抗病毒诊断治疗方面研究的必要性,旨在为培育整体免疫力高的水禽品系奠定理论基础。 相似文献
7.
8.
猪Mx1基因的克隆表达及抗病毒活性研究 总被引:1,自引:0,他引:1
本文应用(RT-PCR)技术,从猪瘟弱毒疫苗和Poly:IC共刺激7 h的PK15细胞的cDNA中扩增出编码Mx1蛋白的全长基因,并通过引物设计填补了PK(15)细胞系Mx1 cDNA序列中3’端11bp的缺失,成功获得Mx1完整基因的克隆。构建了重组表达质粒pRetroQ-sMx1,转染HEK293T细胞并表达Mx1蛋白,利用微量细胞病变抑制法测定其抗病毒活性。双酶切鉴定和核酸序列测定证实pRetroQ-sMx1真核表达质粒构建成功,转染HEK 293T细胞后,能够检测到绿色荧光,Western blot证实为目的蛋白。微量细胞病变抑制法测定重组蛋白具有一定的抗水疱性口炎病毒及猪繁殖与呼吸综合征病毒的活性。重组Mx1具有一定抗病毒生物功能,为进一步研究重组Mx1蛋白的活性以及Mx1抗病毒药物奠定了基础。 相似文献
9.
为探讨不同遗传背景猪黏病毒耐药蛋白1(myxovirus resistance,Mx1)基因的遗传多态性,本试验采用RT-PCR方法扩增克隆出猪Mx1基因并进行生物信息学分析,采用高分辨率熔解曲线法(HRM)对民猪、长白山野猪与民猪杂交猪(简称野杂猪)和大白猪3个群体Mx1基因外显子2区多态性进行分析。结果显示,从民猪及野杂猪中成功克隆出4条Mx1基因mRNA序列。该序列含有1个1 992 bp的完整开放阅读框(ORF),编码663个氨基酸。比较发现4条Mx1基因序列中共含有30个SNPs位点,其中17个SNPs位点引起了氨基酸序列的变异。功能结构域分析发现,大部分氨基酸突变位点位于Mx1基因功能结构域两侧。HRM多态性分析显示,Mx1基因外显子2区DD基因型作为优势基因型在3个群体中具有最高的基因型频率,其中大白猪中DD基因型频率为100%,野杂猪为62.5%,民猪为56.2%。基因多态性分析还发现该区域存在更多的核苷酸及氨基酸突变位点,表明民猪和野杂猪Mx1基因存在更丰富的基因多态性。 相似文献
10.
Mx蛋白为GTP酶动态蛋白家族成员,已被证明具有抗禽流感病毒、Thogoto病毒和水泡性口膜炎病毒等的作用.为制备鸡Mx蛋白的单克隆抗体(MAb),本研究以原核表达并纯化的鸡Mx蛋白为免疫原,免疫BALB/c小鼠,分离脾细胞与SP2/0细胞融合,采用间接ELISA方法筛选阳性克隆,经3次细胞克隆纯化后获得4株稳定分泌抗鸡Mx蛋白抗体的杂交瘤细胞株.间接ELISA测定4株MAb纯化后腹水的效价介于1∶3.2×104~1∶2.5×105之间.Westem blot结果证实4株MAb均具有良好的反应性;间接免疫荧光试验表明其中一株MAb与真核表达的鸡Mx蛋白发生反应.本研究为鸡Mx蛋白功能及表达鸡Mx蛋白的转基因动物等研究奠定了基础. 相似文献
11.
Polymorphisms and the antiviral property of porcine Mx1 protein 总被引:9,自引:0,他引:9
Asano A Ko JH Morozumi T Hamashima N Watanabe T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(12):1085-1089
We determined the cDNA sequences of the type I interferon-inducible proteins, pig Mx1 from PK(15) and LLC-PK1 cells, and compared the antiviral activities of both Mx proteins, including Mx1 polymorphisms against vesicular stomatitis virus (VSV). Mx1 cDNA derived from PK(15) cells had an 11 bp-deletion in the 3' end of the coding region, and was estimated to encode 8 amino acid substitutions and a 23 amino acid extension compared to that from LLC-PK1 cells. VSV replication was inhibited in the 3T3 cells expressing Mx1 mRNA after the cDNA was transfected. However, the efficiency of this inhibition was not different between the cells expressing Mx1 mRNA from both PK and LLC. These results indicate that pig Mx1 protein confers resistance to VSV. 相似文献
12.
13.
Expression of Mx protein and interferon-alpha in pigs experimentally infected with swine influenza virus 总被引:4,自引:0,他引:4
Expression of Mx protein and interferon-alpha (IFN-alpha) was examined by immunohistochemistry in pigs experimentally infected with swine influenza virus. In infected pigs euthanatized at 1 day postinoculation (dpi), the lumen of bronchioles were filled with large numbers of mononuclear cells, small numbers of neutrophils, sloughing epithelial cells, and proteinaceous fluid. Lesions at 3 and 5 dpi were similar but less severe. Alveolar spaces were filled with neutrophils. By 7 and 10 dpi, microscopic lesions were resolved. The immunohistochemical signals for Mx protein and IFN-alpha antigen were confined to cells in areas that had hybridization signal for swine influenza virus. In situ hybridization and immunohistochemistry of serial sections of lung indicated that areas containing numerous swine influenza virus RNA-positive cells also have numerous Mx and IFN-alpha antigen-positive cells. Mean immunohistochemical scores for Mx protein-positive cells were correlated with mean immunohistochemical scores for IFN-alpha antigen-positive cells (r(s) = 0.8799, P < 0.05). These results indicated that Mx protein and IFN-alpha antigen were expressed in the lung from pigs experimentally infected with swine influenza virus, but their biological functions remain to be examined. 相似文献
14.
15.
16.
分别将靶向禽流感病毒(AIV)多靶点siRNA和鸡Mx基因克隆到p201慢病毒表达载体,采用鸡β-actin启动子替换p201慢病毒载体CMV启动子构建重组慢病毒载体p201-β-Mx-siRNA。并将其与辅助包装质粒共转染293T细胞,72h后收集细胞上清,实时荧光定量PCR测定重组慢病毒(rLV-MX-siR-NA)与对照组重组慢病毒(rLV-GFP)CT值,两者比较确定rLV-MX-siRNA滴度。rLV-Mx-siRNA以MOI=4感染猪胎儿成纤维细胞(PEF)并传代培养,提取后代细胞RNA检测外源基因转录以及应用间接免疫荧光试验检测鸡Mx蛋白的表达。结果表明,rLV-Mx-siRNA浓缩后滴度与已知滴度的rLV-GFP相等,为2×108 TU/mL,感染PEF效率>95%。感染rLV-Mx-siRNA的细胞传至第5代和第10代检测到外源基因转录以及鸡Mx蛋白的表达。表达鸡Mx基因和靶向AIV多靶点siRNA重组慢病毒滴度的测定与感染效率的分析,其结果为研究抗AIV转基因猪及其抗AIV的体外评价奠定了基础。 相似文献
17.
Tanabe T Shimoda M Soeno T Suzuki M Tajima M Sato H 《Veterinary immunology and immunopathology》2008,126(1-2):20-26
The interferon-stimulated gene 15 (ISG15) is induced by type I interferon (IFN). Recent studies have revealed that like ubiquitin, ISG15 is conjugated with target proteins. In this study, the feline ISG15 (FeISG15) gene was cloned from feline IFNomega (FeIFNomega)-stimulated feline kidney epithelial (CRFK) cells. According to gene sequence results, cDNA was 474bp long and encoded a protein of 157 amino acids. The putative amino acid sequences showed 62.5-72.1% identity with those of other mammalian ISG15s. Similar to human and mouse ISG15, FeISG15 included tandem ubiquitin-like domains; its homology with feline ubiquitin was 36.3-39.5%. The LRLRGG conjugating motif was located only in the carboxyl terminal ubiquitin-like domain. FeISG15 also lacked the carboxyl terminal extension after the LRLRGG motif, which is present in mouse and human ISG15. Recombinant FeISG15 protein was expressed as a His-tagged fusion protein in Escherichia coli and purified by ion-exchange chromatography followed by affinity chromatography. Monoclonal anti-FeISG15 antibodies revealed free FeISG15 and FeISG15 conjugated with target proteins in cells after IFNomega stimulation by Western blotting analysis. Furthermore, mRNA of IFNgamma was detected from peripheral blood mononuclear cells (PBMCs) after stimulation with rFeISG15 extracellularly by RT-PCR. Taken together, these results suggested that FeISG15 had ubiquitin- and cytokine-like activity, as in other species. 相似文献
18.
ZHANG Li-chun LI Zhao-hua LI Meng-shu SUN Fu-liang CAO Yang PIAO Qing-lin JIN Hai-guo ZHANG Shu-min 《中国畜牧兽医》2017,44(4):1046-1053
In order to explore the polymorphisms of myxovirus resistance (Mx1) gene in pig populations from different genetic background,we cloned Mx1 gene and tested the polymorphisms on exon 2 of Mx1 gene in Min pig, crossbred from wild boars in Changbai Mountain with Min pig (crossbred wild boars) and Large White pig population using high resolution melting (HRM). The results showed that four Mx1 mRNAs were cloned from Min pig and crossbred wild boars, and they had whole ORF which was 1 992 bp in length and coded 663 amino acids. In the four Mx1 mRNA, there were 30 SNPs in nucleotide and 17 mutations in amino acid. The location of SNPs displayed that almost mutations located on two sides of the function domains in Mx1 protein. The polymorphism data in HRM test showed that the DD genotype was the dominant genotype which had the highest frequency in three populations. The DD genotype frequency was 100% in Large White pig population, 62.5% in crossbred wild boars, and 56.2% in Min pig.Some novel mutations in both nucleotide and amino acid on exon 2 region were explored in polymorphism test. In conclusion, there was more polymorphism of Mx1 gene in Min pig and crossbred wild boars. 相似文献
19.
Bracklein T Theise S Metzler A Spiess BM Richter M 《American journal of veterinary research》2006,67(6):1025-1032
OBJECTIVE: To assess the biological response to recombinant feline interferon-omega (rFeIFN-omega) following ocular or oral administration in cats via estimation of Mx protein expression in conjunctival cells (CCs) and WBCs. ANIMALS: 10 specific pathogen-free cats. PROCEDURES: In multiple single-dose drug experiments, each cat received various concentrations of rFeIFN-omega administered topically into both eyes (50 to 10,000 U/eye) and orally (200 to 20,000 units). The same cats received saline (0.9% NaCl) solution topically and orally as control treatments. The CCs and WBCs were collected prior to treatment (day 0), on day 1, and every third or seventh day thereafter until samples yielded negative results for Mx protein. Samples were examined for Mx protein expression via immunohistochemistry and immunoblotting procedures involving murine anti-Mx protein monoclonal antibody M143. RESULTS: After topical application of 10,000 U of rFeIFN-omega/eye, CCs stained for Mx protein for a minimum of 7 days, whereas WBCs were positive for Mx protein for a minimum of 31 days. After topical application of lower concentrations, CCs did not express Mx protein, in contrast to WBCs, which stained for Mx protein at 1,000 units for at least 1 day. Following oral administration, Mx protein was expressed in WBCs at rFeIFN-omega concentrations as low as 200 units, whereas CCs did not stain for Mx protein at any concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that Mx protein expression (a marker of the biological response to rFeIFN-omega) in CCs and WBCs of rFeIFN-omega-treated cats depends on the dose of rFeIFN-omega, site of administration, and cell type. 相似文献