甲基对硫磷人工抗原的合成与鉴定

根据甲基对硫磷的结构特征,分别从甲基对硫磷的硝基一端或磷酸酯基甲氧基一端引入一定长度的间隔臂和活性基团,设计并合成了4种结构的半抗原;针对各种半抗原所带活性基团的性质,通过多种方法制备了与不同的载体蛋白偶联的人工抗原。进行动物免疫后,得到了针对甲基对硫磷的高效价抗体。     

大豆抗原蛋白对幼龄动物致敏作用的研究进展

大豆抗原蛋白对幼龄动物的致敏作用及不易灭活性使其成为大豆及其制品在动物养殖中有效利用的主要瓶颈。本文从主要抗原蛋白的特性、致敏机制、对幼龄动物的影响、降低抗原蛋白的方法及有待进一步探索问题等方面进行了阐述,旨在为高效利用大豆及其制品,以及进一步深入研究提供参考。     

美洲型与欧洲型PRRSV特异N蛋白抗原表位基因表达及其产物的纯化

对原核中以包涵体形式高效表达的美洲型与欧洲型PRRSV特异N蛋白抗原表位区段进行了变性、复性与纯化研究。将美洲型PRRSV重组质粒pGEX-6P-1-N转化入E.coliBL21(DE3)细胞中,成功表达了重组蛋白GST-N,超声裂解复性后目的蛋白经GSTrap FF纯化试剂盒纯化后,纯度可达90%以上。将欧洲型PRRSV重组质粒PQE30-N转化入E.coliM15细胞中表达了重组蛋白His-N。超声裂解后目的蛋白经HisTrapHP蛋白纯化试剂盒纯化后,纯度可达95%以上。Western blot结果表明,两种蛋白分别被美洲型和欧洲型PRRSV阳性血清识别。纯化出的大量美洲型与欧洲型PRRSV特异N蛋白抗原表位区段,为PRRSV的鉴别诊断提供了抗原基础。     

实验感染肉鸡组织中J亚群禽白血病病毒的抗原定位

从实验感染J亚群禽白血病病毒(ALVJ)的髓细胞性白血病肉鸡病例中,选取典型病例,用J亚群禽白血病病毒单克隆抗体通过免疫酶组化试验对不同组织进行了抗原定位观察。结果显示:在实验感染阳性肉鸡体内实质组织细胞及瘤细胞的核膜和细胞浆里可见不同程度的棕黄色阳性反应,可为探讨ALVJ对体内组织细胞的嗜性提供依据。     

Corynebacterium pyrogenes; a biochemical and serological study

The study comprises 136 strains of Corynebacterium pyogenes originating from cattle (105), swine (20), sheep (1), and insects (10). For comparison 2 strains of human origin and 1 strain of Gorynebacterium hemolyticum were examined.One of the bovine strains was atypical, being gelatinase-negative, otherwise the strains of Cb. pyogenes were found to be biochemically identical apart from minor deviations in fermentation patterns (Table 1). Neither were antigenic differences demonstrated (gel diffusion analyses, Figs. 1 and 2).Both of the human strains agreed biochemically with Cb. pyogenes (Table 1). By gel diffusion cross analyses one of them was found to be identical with Cb. pyogenes, the other not, though anti-genically related to it (Fig. 2).Gb. hemolyticum deviated biochemically as well as serologically from Gb. pyogenes, but the 2 organisms shared antigenic determinants (Fig. 3).     

伊氏锥虫在小鼠体内抗原变异规律及免疫保护试验

为了探索伊氏锥虫抗原的变异规律及其用于免疫预防的可能性，首先对伊氏锥虫安微株单虫克隆2个早期变异体ShTatl.1、ShTat1.2采用蛋白酶抑制剂TLCK处理，分离纯化这2种变异体的VSG，用ShTat1.1 VSG免疫KM小鼠，ShTat1.1锥虫攻击免疫鼠后6d分离锥虫，经间接免疫荧光试验（IFT）和班点酶标记试验（Dot-EIA)鉴定为ShTat1.2，说明同一克隆锥虫感染兔，小鼠第1次发生抗原变异后产生的变异体相同。依据这一规律，设计了免疫预防保护试验，试验小鼠分3组：一组为未免疫对照组，一组为ShTat1.1VSG单一抗原免疫组，另一组为ShTat1.1 VSG,ShTat1.2 VSG混合免疫抗原免疫组，各组均以ShTat1.1锥虫攻击，结果ShTat1.1 VSG ShTat1.2VSG免疫组小鼠全部获得保护，单用ShTat1.1 VSG免疫组小鼠和未免疫小鼠血中全部出虫、死亡，前者比后者出虫时间、存活时间延长。提示运用伊氏锥虫抗原变异这一规律设计的这种复合抗原，能激发宿主克服虫体抗原变异对免疫保护的干扰。     

猪细小病毒VP2蛋白主要抗原域在毕赤酵母中的表达及鉴定

猪细小病毒(Porcine parvovirus,PPV)是一自主型细小病毒(Autonomous parvovirus),它是引起母猪繁殖障碍的主要因素之一。目前国内外的诊断方法多以全病毒为抗原,存在很大的局限性,而重组蛋白无感染性,易于生产和纯化。因此用重组蛋白来检测猪细小病中抗体水平对于猪细小病的防     

Genomic, protein and antigenic variability of mycoplasma bovis

Restriction endonuclease analysis (REA) with three enzymes SmaI, PstI, BamHI- was used to identify 13 different genomic groups among 37 Mycoplasma bovis strains. One genomic group was comprised of 14 strains. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) patterns for one strain chosen from each genomic group and an international reference strain PG45 were all similar. Antigenic variability in M. bovis species was investigated by immunoblotting, using serum from a calf that had been naturally infected with M. bovis and three M. bovis-specific monoclonal antibodies — mAbs N₂, I₂ and 5D7. Twenty M. Bovis field strains were tested, comprising one from each genomic group, six from the same genomic group and the reference strain. Antigenic profiles obtained with calf serum differed markedly one from the other, the heterogeneity being equally great among the strains belonging to the same genomic group as those coming from different groups. A stable antigen common to 164 out of 168 strains was detected by mAb N₂, whilst with mAbs I₂ and 5D7, two different membrane antigenic systems were demonstrated that were strikingly variable. These variations in expression occurred not only from one strain to another, but also within the same lineage of clones from a single cell.     

猪传染性胃肠炎病毒NSP8蛋白单克隆抗体的制备及抗原表位的初步鉴定

为鉴定猪传染性胃肠炎病毒(porcine transmissible gastroenteritis virus,TGEV)NSP8蛋白的抗原表位,本研究对GST-NSP8重组蛋白进行了原核表达,并用纯化后的重组蛋白免疫6周龄BALB/c小鼠,取免疫小鼠脾脏,采用常规杂交瘤细胞融合方法,经3次亚克隆后制备了1株稳定分泌抗NSP8蛋白的单克隆抗体杂交瘤细胞株。分泌的单克隆抗体亚类鉴定其重链为IgG1型,轻链为κ链;杂交瘤细胞培养上清的效价为1∶3 200。Western blotting试验结果表明该单克隆抗体能识别原核及真核表达的NSP8重组蛋白。利用截短表达的方法对NSP8蛋白进行抗原表位的鉴定,初步确定了单克隆抗体针对的抗原表位序列为31SPQILKQLTKAFNIAKSDFEREASV55。本研究制备的单克隆抗体及对抗原表位的鉴定,为TGEV NSP8蛋白相关功能的研究奠定了基础。     

克隆伊氏锥虫不同VAT的某些生物学特性研究

对伊氏锥虫安徽株单虫克隆ＳｈＴａｔｌ感染兔体后３、６、９和１８ｄ所分离的４个抗原变异体ＳｈＴａｔｌ．１、ＳｈＴａｔｌ．２、ＳｈＴａｔｌ．３和ＳｈＴａｔｌ．５的生长特征、对人血清敏感性和对小白鼠致病力三个方面进行了比较研究。结果表明不同抗原变异体的生物学特性不完全一致,它提示伊氏锥虫的抗原变异可能伴有虫体其它生理生化特性的改变。