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珍珠鸡是一类具有天然抗病能力的禽种。珍珠鸡对传染性支气管炎、火鸡黑头病、传染性喉气管炎、鸡马立克氏病等有较强的抵抗力[1~ 3] ,珍珠鸡可发生新城疫、鸡白痢、传染病法氏囊炎 ,曲霉菌病[1,4~ 6 ] ,但未能对珍珠鸡的抗病力作明确验证。本试验以珍珠鸡胚为试验对象 ,用新城疫病毒(NDV)、传染病法氏囊炎病毒 (IBDV)、传染性支气管炎 (IBV)及减蛋综合征病毒 (EDSV)来评价珍珠鸡胚胎期对病毒感染的抵抗力。1 材料与方法1 1 毒株 NDV、IBDV、IBV及EDSV均为南京农业大学动物医学院传染病组分离鉴定并保存… 相似文献
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以鸡传染性法氏囊病病毒内蒙古毒株(IBDV-NM)dsRNA为模板,用RT-PCR法扩增其主要寄主保护抗原VP2基因的全长cDNA,克隆于pUC19的XbaI/KpnI位点,进行了全序列分析。序列比较发现IBDV-NM毒株VP2基因与已报道的其它7个IBDVCJ801bkf、Cu1、PBG98、52/70、002—73、STC及VariantE毒株之间高度同源,其核苷酸序列的同源率为91.6%~96.2%,推测的氨基酸序列的同源率为96.2%~98.6%。IBDV-NM毒株VP2高变异区的第一个亲水区氨基酸序列与CJ801bkf、Cu1、PBG98、52/70、STC、002—73比较,有一个氨基酸差异,第二个亲水区氨基酸序列与上述6个毒株完全相同。而与VariantE比较,两个亲水区内各有两个氨基酸差异。此外,IBDV-NM毒株VP2具有强毒株所特有的7肽保守区:SWSASGS。这些结果表明,IBDV-NM毒株为标准血清Ⅰ型IBDV强毒株。 相似文献
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紫外光B辐射增强对水稻叶片内IAA和ABA含量的影响 总被引:9,自引:0,他引:9
两个水稻品种CK46和Dular生长在人工气候箱的条件下,在0.0和13.0kJm^-2day^-1下进行4周的照射处理,研究UV-8对水稻体内内源IAA和ABA含量的影响。结果表明:随着UV-B处理时间的延长,CK46和Dular叶片内的IAA含量下降。相反,UV-B辐射增强使两个品种叶片内的ABA含量上升。 相似文献
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马立克氏病病毒(MDV)814株B抗原基因的克隆及部分序列分析 总被引:6,自引:1,他引:6
提取感染鸡胚成纤维细胞MDV-I弱毒株814病毒DNA为模板,根据RBIB株gB基因5′及3′两端核苷酸离列设计引物,利用PCR技术扩增了我国MDV-I弱毒株814gB基因(2.9kb)将扩增片段平末端克隆到载体pBluescriptSK中EcoRV位点,经BamHI,HindIII酶切鉴定得到不同插入方向的重组质粒。构建圹增片段的酶切图谱及部分序列分析证明与RBIB株gB基因无差异,显示了极高的 相似文献
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以荧光诱导动力学、低温荧光发射光谱及希尔反应活性测定为手段,研究了小麦、大豆等高等植物风干叶片(相对含水量(1.5±0.2)%)及其叶绿体的光合活性。暗中风干并保存的叶片照光时仍能进行电荷分离、QA还原及至少包含QA-→QB在内的次级电子传递,其复水后的叶绿体具有光推动的光系统I电子传递活性(DCIPH2→MV)和包含光解水放氧能力在内的光系统II电子传递活性(H2O→DMBQ),但没有全光合链(H2O→MV)的电子传递活性。光系统II核心天线的77K荧光峰位(686nm和694nm)不受脱水的影响,而光系统I外周天线LHCI的77K荧光峰位对脱水十分敏感,叶片风干过程中从739nm移到726nm。这些结果表明,光合作用器中越靠近反应中心核心的组分的组织结构越紧密有序,其结构和功能越少受快速水胁迫的影响;在整个光合电子链中,受快速水胁迫影响最大的部位在两个光系统之间,这个部位的电子传递被风干处理不可逆地阻断。 相似文献
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牛病毒性腹泻病毒(BVDV)对牛免疫缺陷病毒(BIV)的激活作用 总被引:6,自引:0,他引:6
通过合胞体分析和反转录酶活力测定,首次证明牛病毒性腹泻病毒能激活牛免疫缺陷病毒的复制与表达,并进一步通过转染实验和凝胶电泳漂移分析证明,当BIV LTR的NF-k B区缺失时,BVDV则不能实现其激活作用,BVDV直接或间接诱导牛NF-kB因子作用于BIV LTR的NF-kB区实现其激活作用。 相似文献
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鸡传染性支气管炎病毒中国流行株免疫原S1基因的分子克隆、序列分析及其DNA免疫的初步研究 总被引:1,自引:0,他引:1
对来源于我国华东地区的鸡传染性支气管炎病毒流行株QD免疫原S1基因cDNA进行了克隆、序列分析和DNA免疫的初步研究。RTPCR扩增QD毒株的S1基因,将其5′和3′端分别进行分子修饰后插入克隆载体pUC18的BamHⅠ/HindⅢ位点,在大肠杆菌中实现了目的基因的克隆;利用英国IBV毒株S1全基因核酸探针与QD毒株S1基因的重组克隆质粒分子杂交后,采用HaeⅢ,PvuⅡ和XbaⅠ等限制酶对此流行毒株S1基因cDNA进行了酶切分析;在测定QD毒株S1基因5′端高变区核苷酸序列并以此与IBVM41,H120,6/82及Beaud等参考毒株序列对比分析的基础上,构建了QD株S1基因DNA免疫表达质粒,肌肉注射免疫小鼠后,鸡胚病毒中和试验的结果表明,IBVS1基因DNA免疫表达质粒能诱导小鼠产生病毒特异的中和抗体,具有良好的免疫原性,初步显示基因疫苗在鸡传染性支气管炎防治上应用前景。 相似文献
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目前传染性法氏囊病病毒疫苗主要是用原代鸡胚成纤维细胞 (PCEF)增殖IBDV进行生产。由于SPF种蛋价格高 ,且SPF种蛋在取得及培养过程中易被外源病原污染 ,造成产品质量的不稳定 ,生产成本很高[1] 。Vero细胞系是一种贴壁依赖性的传代细胞系 ,WHO已经批准用Vero细胞作为载体进行病毒疫苗的生产。目前已成功地应用Vero细胞生产出脊髓灰质炎病毒疫苗和狂犬病毒疫苗[2 ] 。用Vero细胞生产传染性法氏囊病病毒疫苗也会有较好的前景。我们已完成了在Vero细胞上静止状态下增殖IBDV弱毒株的培养条件研究 ,而… 相似文献
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S M Tian K C Ruan J F Qian G Q Shao C Balny 《European journal of biochemistry》2000,267(14):4486-4494
The effects of high hydrostatic pressure on the structure and biological activity of infectious bursal disease virus (IBDV), a commercially important pathogen of chickens, were investigated. IBDV was completely dissociated into subunits at a pressure of 240 MPa and 0 degrees C revealed by the change in intrinsic fluorescence spectrum and light scattering. The dissociation of IBDV showed abnormal concentration dependence as observed for some other viruses. Electron microscopy study showed that morphology of IBDV had an obvious change after pressure treatment at 0 degrees C. It was found that elevating pressure destroyed the infectivity of IBDV, and a completely pressure-inactivated IBDV could be obtained under proper conditions. The pressure-inactivated IBDV retained the original immunogenic properties and could elicit high titers of virus neutralizing antibodies. These results indicate that hydrostatic pressure provides a potential physical means to prepare antiviral vaccine. 相似文献
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摘要:【目的】本研究旨在构建在鸡原代骨骼肌细胞中表达IBDV病毒VP2基因的重组杆状病毒。【方法】从IBDV适应细胞毒中提取RNA,用RT-PCR技术扩增VP2基因,将其克隆到自主构建的杆状病毒转移载体的CMV启动子之下,通过Bac-to-Bac系统获得VP2重组Bacmid,并将其转染Sf9昆虫 细胞,获得了VP2重组杆状病毒。重组病毒经扩增后以50个MOI感染鸡原代骨骼肌细胞,接种72h后裂解细胞收获蛋白。【结果】蛋白样品经SDS-PAGE和Western blot证实VP2蛋白获得表达,分子量约48kDa,与预测蛋白大小一致,且能被IBDV阳性血清所识别。【结论】重组杆状病毒可以有效地将VP2基因导入鸡原代细胞,并在CMV的启动下表达具有抗原性的VP2蛋白,本研究为研制IBDV及其他重要禽类传染病的杆状病毒载体疫苗奠定了基础。 相似文献
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Feng Chen Jiajia Liu Zhuanqiang Yan Di Liu Jun Ji Jianping Qin Haiyan Li Jingyun Ma Yingzuo Bi Qingmei Xie 《Journal of virology》2012,86(21):11942-11943
A novel isolate of infectious bursal disease virus (IBDV) was designated GX-NN-L. The GX-NN-L IBDV was a very virulent infectious bursal disease virus (vvIBDV) isolated from broiler flocks in Guangxi province, China, in 2011. The GX-NN-L IBDV caused high mortality, immunosuppression, low weight gain, and bursal atrophy in commercial broilers. Here, we report the complete genome sequence of the GX-NN-L IBDV, a reassortment strain with segments A and B derived from very virulent strains and attenuated IBDV, respectively. These findings from this study provide additional insights into the genetic exchange between attenuated and very virulent strains of IBDV and continuous monitoring of the spread of the virus in chicken. 相似文献
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Wei Y Li J Zheng J Xu H Li L Yu L 《Biochemical and biophysical research communications》2006,350(2):277-287
Infectious bursal disease virus (IBDV), a double-stranded RNA virus, is a member of the Birnaviridae family. Four pathotypes of IBDV, attenuated, virulent, antigenic variant, and very virulent (vvIBDV), have been identified. We isolated and characterized the genomic reassortant IBDV strain ZJ2000 from severe field outbreaks in commercial flocks. Full-length genomic sequence analysis showed that ZJ2000 is a natural genetic reassortant virus with segments A and B derived from attenuated and very virulent strains of IBDV, respectively. ZJ2000 exhibited delayed replication kinetics as compared to attenuated strains. However, ZJ2000 was pathogenic to specific pathogen free (SPF) chickens and chicken embryos. Similar to a standard virulent IBDV strain, ZJ2000 caused 26.7% mortality, 100% morbidity, and severe bursal lesions at both gross and histopathological levels. Taken together, our data provide direct evidence for genetic reassortment of IBDV in nature, which may play an important role in the evolution, virulence, and host range of IBDV. Our data also suggest that VP2 is not the sole determinant of IBDV virulence, and that the RNA-dependent RNA polymerase protein, VP1, may play an important role in IBDV virulence. The discovery of reassortant viruses in nature suggests an additional risk of using live IBDV vaccines, which could act as genetic donors for genome reassortment. 相似文献
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传染性法氏囊病病毒在次代鸡胚成纤维细胞上的增殖 总被引:2,自引:0,他引:2
研究了用次代鸡胚成纤维细胞(SCEF)增殖传染性法氏囊病病毒(IBDV)的可能性。在研究了原代鸡胚成纤维细胞(PCEF)与SCEF的生长特性的基础上,就各种培养方式采用PCEF与SCEF增殖IBDV进行了比较。结果表明,可用SCEF代替PCEF进行IBDV的增殖培养。 相似文献
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A recombinant Newcastle disease virus (NDV) expressing VP2 protein of infectious bursal disease virus (IBDV) protects against NDV and IBDV 总被引:18,自引:0,他引:18
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Infectious bursal disease virus (IBDV) causes a highly immunosuppressive disease in chickens. Currently available, live IBDV vaccines can lead to generation of variant viruses. We have developed an alternative vaccine that will not create variant IBDV. By using the reverse genetics approach, we devised a recombinant Newcastle disease virus (NDV) vector from a commonly used vaccine strain LaSota to express the host-protective immunogen VP2 of a variant IBDV strain GLS-5. The gene encoding the VP2 protein of the IBDV was inserted into the most 3'-proximal locus of a full-length NDV cDNA for high-level expression. We successfully recovered the recombinant virus, rLaSota/VP2. The rLaSota/VP2 was genetically stable, at least up to 12 serial passages in chicken embryos, and was shown to express the VP2 protein. The VP2 protein was not incorporated into the virions of recombinant virus. Recombinant rLaSota/VP2 replicated to a titer similar to that of parental NDV strain LaSota in chicken embryos and cell cultures. To assess protective efficacy of the rLaSota/VP2, 2-day-old specific-pathogen-free chickens were vaccinated with the recombinant virus and challenged with a highly virulent NDV strain Texas GB or IBDV variant strain GLS-5 at 3 weeks postvaccination. Vaccination with rLaSota/VP2 generated antibody responses against both NDV and IBDV and provided 90% protection against NDV and IBDV. Booster immunization induced higher levels of antibody responses against both NDV and IBDV and conferred complete protection against both viruses. These results indicate that the recombinant NDV can be used as a vaccine vector for other avian pathogens. 相似文献
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Zhang L Hu TJ Liu HL Shuai XH 《International journal of biological macromolecules》2011,49(4):607-615
The aim of this study was to investigate the inhibitory effect of Sargassum polysaccharide on oxidative stress induced by infectious bursa disease virus (IBDV) in chicken bursal lymphocytes. The levels of IL-1β, IL-8, IL-10, TNF-α, MCP-1, reduced glutathione and reactive oxygen species in chicken bursal lymphocytes treated with IBDV or both IBDV and Sargassum polysaccharide were measured, and the activities of superoxide dimutase and glutathione peroxidase were evaluated. Our results showed that oxidative stress appeared when chicken bursal lymphocytes were incubated with IBDV for 8 h at 100 TCID50. Sargassum polysaccharide inhibited oxidative stress by increasing the amount of reduced glutathione, promoting the activities of superoxide dimutase and glutathione peroxidase and reducing the level of reactive oxygen species. The polysaccharide also raised IL-1β, IL-8, IL-10 and TNF-α levels in cells infected with IBDV. These findings suggest that Sargassum polysaccharide acts against infection by elevating antioxidant capacity and cytokine levels in chicken bursal lymphocytes. 相似文献
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应用重叠延伸剪切技术(splicing by overlapping extension,SOE),经3次PCR将传染性法氏囊病病毒(infectiousbursal disease virus,IBDV)多聚蛋白(VP2/4/3)基因和鸡白细胞介素2(Chicken IL-2,ChIL-2)基因进行融合,定向插入真核表达载体pCI的CMV启动子下游,获得重组质粒pCI-VP2/4/3-IL-2和pCI-IL-2-VP2/4/3。将其制备成DNA疫苗,肌肉注射14日龄非免疫鸡,2周后加强免疫,定期测定鸡抗IBDV血清ELISA抗体效价及病毒中和抗体效价。加强免疫后3周用IBDV标准强毒株攻击,连续观察3天后全部扑杀,计算保护率及囊体比,并进行组织病理学检查。结果表明:1)融合基因重组质粒pCI-VP2/4/3-IL-2、pCI-IL-2-VP2/4/3免疫后能明显增强IBDVDNA疫苗对强毒的攻击保护(保护率分别为83.3%、91.6%),显著高于pCI-VP2/4/3单独免疫对照组(58.3%);2)诱导产生的抗IBDV血清ELISA抗体效价明显增高(P<0.05),同时能提高DNA疫苗诱导产生的中和抗体效价(P<0.05);3)能显著促进鸡外周血液T淋巴细胞增殖反应。上述结果提示:IBDV VP2/4/3与ChIL-2基因融合后发挥了相互协同作用,ChIL-2产生了分子免疫佐剂效应;融合基因DNA疫苗能增强IBDV DNA疫苗的免疫原性,促进了机体特异性免疫应答。 相似文献
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Viruses induce dramatic changes in target tissue during pathogenesis, including host cellular responses that either limit or support the pathogen. The infectious bursal disease virus (IBDV) targets primarily the bursa of Fabricius (BF) of chickens, causing severe immunodeficiency. Here, we characterized the cellular proteome changes of the BF caused by IBDV replication in vivo using 2DE followed MALDI-TOF MS identification. Comparative analysis of multiple 2DE gels revealed that the majority of protein expression changes appeared between 24 and 96 h after IBDV infection. MS identified 54 altered cell proteins, 12 of which were notably upregulated by IBDV infection. Meanwhile, the other 42 cellular proteins were considerably suppressed by IBDV infection and are involved in protein degradation, energy metabolism, stress response, host macromolecular biosynthesis, and transport process. The upregulation of β-actin and downregulation of dynamin during IBDV infection were also confirmed by Western blot and immunofluorescence analysis. These altered protein expressions provide a response profile of chicken BF to virulent IBDV infection. Further functional study on these altered proteins may lead to better understanding of pathogenic mechanisms of virulent IBDV infection and to new potential therapeutic targets. 相似文献
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鸡胚成纤维细胞cDNA表达文库的构建 总被引:2,自引:0,他引:2
鸡胚成纤维细胞(CEF)是研究鸡传染性法氏囊病病毒(IBDV)的主要细胞材料,而构建CEF的cDNA表达文库是筛选IBDV在CEF中的细胞受体,研究细胞嗜性的基础平台。采用Gateway技术构建CEF的表达文库,避免使用限制性内切酶切割cDNA,能够解决常规方法构建cDNA文库的技术缺陷。该技术将CEF的mRNA分离纯化后,以5′端生物素标记的Oligo(dT)primer为引物反转录后连接Adapter,层析柱纯化,通过BP重组反应构建cDNA入门文库,其平均滴度为1.1×106cfu/mL,文库总容量为1.2×107cfu,平均插入片段为2243bp,重组率为100%。通过LR重组反应将入门文库转换为表达文库,经测定平均滴度为5×105cfu/mL,文库总容量为5.5×106cfu,平均插入片段为2411bp,重组率为100%。结果表明,所构建的文库具有较高的重组率和较大的库容量,可作为较高质量的文库来研究IBDV的相关基因,为研究病毒受体和病毒入侵途径,进一步了解IBDV的致病机理奠定了基础。 相似文献