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1.
草鱼出血病病毒多肽的基因定位   总被引:15,自引:3,他引:12  
用聚丙烯酰胺凝胶电泳分离纯化的草鱼出血病病毒GCHV873的基因组ds-RNA的11个片段,分别在麦胚无细胞翻译体系中进行翻译。其翻译产物经SDS-PAGE系统分析。结果表明,基因组片段1、2、3、4、5、和10分别编码病毒核心衣壳的结构多肽VP1、VP2、VP3、VP4、VP5和VP10,片段6和7分别编码病毒外层衣壳的结构多肽VP6和VP7。片段8和9分别编码52kD和41kD的多肽,片段11编码两种多肽,分子量分别为29kD和19.5kD,它们与病毒结构多肽无明显对应关系。病毒基因组与多肽大体是一一对应的关系。  相似文献   

2.
黑龙江省及长春市烟草病毒病的种类鉴定   总被引:4,自引:0,他引:4  
1991-1993年在黑龙江省主要烟区的11个县及长春市采样,得129个毒株。经鉴别寄主测定,抗血清反应(板式酶联法或斑点酶联法)及电镜或免疫电镜观察,有TMV(43.4%),PVY(17.8%),CMV(3.9%),TRV(0.8%),TSWV等病毒,TMV与PVY混合侵染的占10.1%,PVY与其它病毒混合侵染的占11.6%,另有5个标样为马铃薯Y病毒组成员,10个为未知。  相似文献   

3.
人类疱疹病毒6型感染细胞免疫学特性研究   总被引:2,自引:0,他引:2  
赵蓓  姚坤 《Virologica Sinica》1998,13(3):232-236
采用间接免疫荧光法、APAAP法及MTT法,研究了人类疱疹病毒6型(HHV6)中国南京地方株CN5感染细胞病毒抗原表达的形态学和动力学特征、CD抗原表达阳性细胞百分率的变化及PHA诱导的细胞增殖反应的改变。结果显示,CN5感染脐血单个核细胞(CBMCs)后8~12h即可在细胞内检出病毒抗原,至接种后48h,病毒抗原阳性细胞可达36%;CN5感染CBMCs和成人外周血单个核细胞(PBMCs)后可引起两者CD3阳性细胞减少、CD4阳性细胞增多,而对CD2、CD8、CD45RA阳性细胞百分率未见明显影响;CN5感染细胞裂解液对PHA诱导的PBMCs增殖反应具有抑制作用,这种抑制作用与该裂解液的蛋白浓度之间呈一剂量依赖关系,且可被HHV6抗血清所逆转。  相似文献   

4.
兔出血症病毒结构多肽分析   总被引:3,自引:0,他引:3  
粗提病毒经Sepharose4B层析后,获得纯化的兔出血症病毒(RHDV)。提纯病毒经过SDS-PAGE经考马斯亮蓝染色显示A、B、C、D、E、F和G7条多肽,凝胶扫描显示A为RHDV主要结构多肽。用多抗和单抗作免疫转印分析,证实A、B、C、D、E、和G为结构多肽,此6条结构多肽间的抗原关系十分密切。  相似文献   

5.
应用差别p H 值沉淀蛋白质的原理,建立了水稻条纹病毒病特异蛋白( S P) 的两种提纯方法。这两种方法都可以从病叶中提纯到大量的 S P,其粗提纯量分别为0 .8 和2 .0 mg/g 病叶。通过 S D S P A G E 分离后得到了精提纯的蛋白,其分子量为20 .1 k Da 。将粗提纯和精提纯的 S P 分别免疫兔子,制备出效价为51 200 和6 400 的抗血清。将效价为6 400 的高度特异性的抗血清用于研究 R S V S P 与 R S V C P 及同属的水稻草状矮化病毒( R G S V) S P、 C P 之间的血清学关系,结果表明, R S V S P 的抗血清与 R G S V C P、 R S V C P 之间无反应,但可与 R G S V S P 微弱反应;而 R G S V S P、 C P 及 R S V C P 的抗血清与 R S V S P 之间都无血清学反应。结果证实了 R S V 和 R G S V 之间存在着进化上的亲缘关系  相似文献   

6.
将马铃薯Y病毒普通系(PVY0)的外壳蛋白基因克隆到表达质粒pMALc2中,构建这一基因在大肠杆菌中的表达载体pMALc2PVY0CP。SDSPAGE及Westernbloting检测结果表明,这一表达栽体在E.coliDH5α中经IPTG诱导可表达分子量为71.8kDa的特异性融合蛋白。以amyloseresin亲合柱层析纯化这一融合蛋白为抗原,免疫家兔制备了效价为1∶1024的特异性抗血清。用该抗血清可通过对流免疫电泳、免疫双扩散及Westernbloting对PVY进行检测  相似文献   

7.
草鱼出血病病毒多肽的荧光染色   总被引:1,自引:0,他引:1  
王炜  陈延 《Virologica Sinica》1994,9(2):157-159
将草鱼出血病病毒(GrassCarpHemorrhageVirus,GCHV)置于还原性的溶液中,然后加入等体积的NaHCO3配制的异硫氰酸荧光索溶液进行多肽的标记,再经SDS-PAGE分析,在紫外灯下即可检测到GCHV全部的11个结构多肽的荧光带。该方法最小检测量为500ng,由该方法回收的多肽具有抗原活性,可作为抗原进行免疫学实验。  相似文献   

8.
高效价甘薯羽状斑驳病毒抗血清的制备   总被引:6,自引:0,他引:6  
用嫁接方法将甘薯羽状斑驳病毒(SPFMV)接种到I.setosa上扩繁,以0.2mol/LpH7.2PBK缓冲液、垫层差速离心、蔗糖密度梯度离心提取纯化SPFMV。纯化的SPFMVOD260/280的比值为1.25。将纯化的SPFMV免疫家兔制备抗血清,在环状沉淀和微量沉淀试验中,用提纯病毒测定抗血清的效价均为1:4096;以SPFMV-IgG为第一抗体,应用Dot-ELISA对甘薯和I.selosa叶片中的SPFMV分别作了测定。  相似文献   

9.
蔗糖密度梯度离心法提纯斜纹夜蛾核多角体病毒($Spodoptera litura  multinucleocapsid nucleopolyhdrovirus, SpltMNPV)包埋型病毒粒子(polyhedra-derived virus,PDV),以此病毒粒子作抗原,免疫家兔获得抗血清;用SDS裂解缓冲液提取斜纹夜蛾幼虫中肠组织细胞总蛋白。采用病毒铺覆蛋白印迹技术(VOPBA),利用抗PVD抗血清对病毒受体进行检测,结果表明斜纹夜蛾中肠细胞总蛋白中40kD、73kD、85kD的三种蛋白能够结合PDV。  相似文献   

10.
腮腺炎病毒抗血清的制备   总被引:3,自引:0,他引:3  
以制备适用于疫苗生产检定中病毒鉴别试验和外源因子检查的高效价腮腺炎病毒抗血清为目的。用腮腺炎病毒接种SPF鸡胚尿囊腔,培养收取病毒尿液免疫SPF鸡,采集抗血清。腮腺炎病毒接种Vero细胞,培养病毒抗原经PEG沉淀,超速离心法纯化后免疫家兔采集抗血清。比较两种免疫方法所得病毒抗血清效价。结果显示SPF鸡抗腮腺炎病毒血清中和抗体GMT为1:1716,兔抗腮腺炎病毒血清中和抗体GMT为1:732。两种动物抗血清均适用于疫苗生产相关检定。免疫SPF鸡制备的病毒抗血清无特定病原及抗体污染,是毒种外源因子检测和疫苗鉴别试验的理想试剂。免疫SPF鸡制备病毒抗血清的程序简单,结果易于验证,有利于生物试剂标准化。  相似文献   

11.
Antibodies to disrupted murine sarcoma-leukemia virus (MSV[MLV]) were used to study the synthesis of viral polypeptides in the transformed, virus-producing rat cell line 78A1. When cultures were labeled for 10 min with radioactive amino acids, about 9% of the total labeled proteins were precipitated with antiserum against purified MSV(MLV), and 3 to 4% were precipitated with the same antiserum after it had been absorbed with an extract from uninfected rat cells. The difference is due to the presence in the unabsorbed antiserum of antibodies to cellular proteins that are present in purified virus preparations. Intracellular viral proteins labeled with radioactive amino acids were isolated by immunoprecipitation and analyzed by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. The mobilities of intracellular viral polypeptides were identical to those of the purified virion. However, labeled polypeptides having electrophoretic mobilities lower than that of the major virion polypeptide, the group-specific antigen of molecular weight 31,000, were present in higher proportion in the total cell extract and in the membrane fraction than in the virion. These polypeptides appear to be of cellular origin for they were present only in minute amounts in the immunoprecipitates obtained with the absorbed serum. After a 10-min labeling period, radioactive proteins were assembled into extracellular virions rapidly for the first 4 hr followed by a slower rate. More than 2% of the total proteins of the cell labeled in a 10-min pulse were assembled into virions at the completion of a 24-hr chase. The high-molecular-weight polypeptides with the same mobilities as those detected in the immunoprecipitate of intracellular proteins were found in virions released from cells after a 10-min pulse. A larger proportion of these high-molecular-weight proteins was detected in virions released after short chase periods (30-120 min) than after longer chase periods (6-24 hr). Two possible interpretations of these data are that the high-molecular-weight cell-derived polypeptides (i) have a turnover rate higher than that of the major virion polypeptides or (ii) are cleaved proteolytically from the virions during long incubation in the culture media.  相似文献   

12.
Serotype-specific monoclonal antibodies were used to select mutants of SA11 rotavirus that were resistant to neutralization. The antigenic characteristics of these mutants were studied with with a panel of monoclonal antibodies. We isolated one type of mutant which showed a dramatic increase (greater than 10-fold) in resistance to neutralization by hyperimmune antiserum, and this together with other data indicates the presence on the rotavirus major outer shell glycoprotein of an immunodominant antigenic site involved in virus neutralization. The mutants were also useful in classifying neutralizing monoclonal antibodies.  相似文献   

13.
The polypeptides of reticuloendotheliosis virus (REV) were separated by gel filtration in the presence of guanidine hydrochloride. The eight peaks obtained by gel filtration were then analyzed by polyacrylamide gel electrophoresis and four appeared to contain single polypeptides. The material identified as p29 was used to prepare antiserum. This protein constitutes the major internal non-glycosylated polypeptide in the virion. Double immunodiffusion indicated that the antiserum was specific for p29. Using this antiserum, cross-reactivity was demonstrated between REV, chick syncytial virus, duck infectious anemia virus, and spleen necrosis virus. Antiserum to p29 failed to cross-react with Rous sarcoma virus. This indicates that p29 is a group-specific antigen shared by the viruses of the REV complex. A microcomplement fixation test was developed with this antiserum that will be useful in the quantitation of REV and the identification of other members of this newly defined group.  相似文献   

14.
The sensitivities of double-immunodiffusion (DID) and neutralization tests to detect avian encephalomyelitis (AE) antibody in chickens were studied. Two antigens were employed in the tests. Concentrated antigen gave a higher titer of antiserum than crude antigen, which reacted only to serum having a neutralization log-index (NI) of 3.4approximately4.0 or more. Antibody responses were examined in four growing chick groups inoculated with AE virus by the intracerebral, subcutaneous and oral routes by the DID test with concentrated antigen and by the neutralization test for 1 or over 2 years after inoculation. When concentrated antigen was used, most sera having an NI of over 1.0 were positive for precipitating antibody. Therefore, the sensitivity of the DID test was nearly equal to that of the neutralization test. The DID test was considered to be applicable to the diagnosis of AE and an antibody survey in the field.  相似文献   

15.
The immunological properties of a glycoprotein antigen (antigen 2) ofStreptococcus agalactiae serotype Ia were investigated. A specific antiserum was prepared by immunizing rabbits with antigen 2 immunoprecipitates excised from Crossed immunoelectrophoresis (Crossed IEP) gels. This antiserum produced a single peak representing antigen 2 when reacted with a Triton X-100 sonicate of heat-killed whole serotype Ia cells in Crossed IEP analysis. With polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and subsequent immunoelectroblotting, three strongly reacting polypeptides were detected at 60, 56, and 35 kilo-Daltons. Many faintly reacting polypeptides were detected between 67 and 30 kilodalton. The specific anti-antigen 2 serum used in Crossed immunoisoelectric focusing (XIEF) detected three immunoprecipitates, two with a pI of 8.4 and one with a pI of 6.7. Identification of the antigens detected in XIEF with the polypeptides detected by immunoelectroblotting was not attempted. The specific anti-antigen 2 serum partially protected mice against lethal serotype Ia infection.  相似文献   

16.
草鱼出血病病毒的糖蛋白和结构多肽的抗原性   总被引:6,自引:0,他引:6  
陈延  王炜 《病毒学报》1992,8(1):57-61
  相似文献   

17.
In this article, the preparation and characterization of polyclonal rabbit antisera against the individual polypeptides of bovine neurofilament (68, 150, and 200 kilodaltons) is described. Selected antisera against the 68- and 150-kilodalton neurofilament polypeptides were specific for the corresponding antigen in homogenates of bovine, rat, and human brain as judged by immunoblots. The antisera against the 200-kilodalton neurofilament polypeptide cross-reacted to some extent with the 150-kilodalton neurofilament polypeptide, especially with the human antigen. The most specific antisera were used to develop an enzyme-linked immunosorbent assay (ELISA), and the cross-reactivities between the antisera and the different bovine and rat neurofilament polypeptides were determined. Contrary to the results in the immunoblots, the antiserum against the 200-kilodalton neurofilament polypeptide was subunit-specific, as was the 150-kilodalton antiserum. The 68-kilodalton antiserum displayed a minute cross-reactivity against bovine 150- and 200-kilodalton neurofilaments, but it cross-reacted somewhat more with the rat 150- and 200-kilodalton antigens. Even so, the subunit specificity of the antisera is high enough to enable the development of a quantitative ELISA for determination of the individual bovine or rat neurofilament polypeptides in a mixture. This study is the necessary preparation for such an assay.  相似文献   

18.
For differentiation of Ilvin-Bykovsky virus (IBV) and monkey Meson-Pfeizer virus (M-PMV) the method of virus neutralization with antibodies against the envelope virus antigen was used. The viruses were cultivated in similar human embryo cells. The results of the virus neutralization were determined by presence or absence of the gs-antigen in the infected cells. The antiserum to M-PMV envelope antigens did not neutralize the IBV antigen. It has been concluded that IBV and M-PMV differ by their envelope antigens and should be regarded as different viruses.  相似文献   

19.
Polyadenylated RNA isolated from total polyribosomes of two variable antigen types (VATs) of T. brucei brucei were shown to program the synthesis, in mRNA-dependant reticulocyte lysates, of a wide variety of polypeptides. After immunoprecipitation of these cell-free products with an homologous antiserum raised against purified variant-specific surface antigen (VSSA), a major electrophoretic band was apparent on fluorography. It was confirmed that this band corresponds to the variable antigen since only an excess of purified homologous antigen will provoke competition. The apparent molecular weight of the in vitro synthesized antigen is about 63,000 daltons. The VSSA mRNA has been found in membrane-bound polyribosomes and a 15 fold immunological purification of this mRNA has been obtained, using partially purified anti-VSSA IgG in conjunction with inactivated Staphylococcus aureus.  相似文献   

20.
The major polypeptides (P-1, P-2, and P-6) of HBsAg were isolated from purified preparations of 22-nm HBsAg particles, iodinated, and analyzed by double-antibody radioimmunoprecipitation assays for the presence of hepatitis B virus (HBV)-specific antigens. Each polypeptide fraction contained both group (a) and subtype (d) specific determinants in common by virtue of their immunoreaction with antiserum to native HBsAg and antisera to the other structural polypeptides. The antigenic and structural similarities of the HBsAg polypeptides establish that they are not each unique gene products of the HBV genome.  相似文献   

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