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1.
利用噬菌体随机7肽库分析猪繁殖与呼吸综合征病毒(PRRSV)N蛋白B细胞抗原表位。以PRRSV BJ-4株N蛋白的单克隆抗体(mAb)GE3作为筛选分子，筛选噬菌体展示的随机7肽库。经序列测定和分子生物学软件分析，表明噬菌体展示的外源氨基酸序列与PRRSV BJ-4株N蛋白的50～55aa的氨基酸序列有极高的同源性。ELISA分析结果表明，筛选到的噬菌体能特异性地与PRRSV阳性血清结合，从而证实了该表位是单克隆抗体GE3所识别的抗原表位。 相似文献
2.
选取2002～2007年从上海市分离鉴定的11株鸡新城疫病毒（Newcastle disease virus,NDV）强毒株,克隆其融合蛋白（Fusion,F）基因片段,测序后登录GenBank,序列号分别为NDV 022433（EU258661）、022435（EU258662）、0210149（EU258653）、0210154（EU258654）、0210227（EU258656）、0210252（EU258658）、03024（EU258637）、04011（EU258639）、05013（EU258640）、07011（EU258642）和07023（EU258643）。序列分析结果显示：上述分离株的F基因扩增片段为1 654 bp,编码551个氨基酸,其核苷酸序列同源性为94.6%～100%,推导的F蛋白氨基酸序列同源性为93.0%～100%;各分离株F蛋白裂解位点序列均为112R-R-Q-K-R-F117,为基因Ⅶ型NDV强毒株。系统发育分析表明：这11株NDV分离株之间的遗传距离较近,而与传统疫苗株B1、La Sota、V4和F48E9的遗传距离较远。此外,这11株NDV强毒株均具有大多数鹅源NDV毒株特有的973和1 249 nt RsaⅠ位点。 相似文献
3.
利用噬菌体随机肽库筛选传染性法氏囊病病毒VP2模拟表位 总被引：2，自引：0，他引：2
用纯化的4株传染性法氏囊病病毒(IBDV)VP2单克隆抗体(1B5,5D1,2H11和I4-4-3)筛选噬菌体随机12肽库,对40个克隆(10个单克隆噬菌斑×4株单克隆抗体)进行ELISA验证,获得32个阳性克隆,选20个阳性克隆(5个单克隆噬菌斑×4株单克隆抗体)进行DNA序列分析,确定了4个优势12肽为不同传染性法氏囊病病毒(IBDV)抗原表位.这4个12肽在一级结构上没有3个以上连续氨基酸与GenBank中传染性法氏囊病病毒、VP2的氨基酸序列相同之处,与单抗的结合可被VP2蛋白有效地抑制,推测可能是构象依赖性表位.将4个模拟表位串联表达后获得目的蛋白,经SDS-PAGE分析,目的蛋白占菌体总蛋白的20%,分子量30 kD.用多克隆抗体对目的蛋白进行免疫印迹分析,结果表明目的蛋白具有特异性和反应原性.试验结果提示:筛选的是传染性法氏囊病病毒(IBDV)VP2单抗的模拟表位. 相似文献
4.
一株信鸽新城疫强毒的分离鉴定和分子特性分析 总被引：9，自引：0，他引：9
从发病鸽的体内分离到一株新城疫病毒（Newcastle Disease Virus，NDV）PB9601，经病毒蚀斑克隆纯化后动物回归试验可产生典型的NDV病变，其MDT、ICPI、IVPI分别为50.5、1.65和2.36，表明为NDV强毒。对其F和HN蛋白分别进行基因克隆测序，发现F蛋白多肽裂解位点的序列为111 KRQKRF 117，符合鸽源NDV强毒氨基酸基序。F基因分型及同源性比较显示：PB9601与YN-03、TW-98等鸽源毒株属于VI型，其F基因与基因Ⅱ型疫苗株LaSota氨基酸同源性为88.6%，与基因Ⅰ型疫苗株V4的氨基酸同源性为91.0%，与基因Ⅸ型传统强毒F48E9等的氨基酸同源性为90.6%；与国内鸽分离株GX-05、YN-03、TW-98的氨基酸同源性分别为92.1%、95.5、98.6%、与国外鸽分离株Capital3-97、Tigre6-99、1166-00、IT-227-82、2736-00的同源性分别为93.3%、91.9%、94.6%、95.1%、93.6%。HN基因氨基酸同源比较显示：PB9601与LaSota、F48E9、V4等的氨基酸同源性分别为87.2%、89.2%、87.6%；与国外鸽分离株1166-00、IT-227-82、2736-00的同源性为93.7%、95.5%、92.3%。由此看出PB9601与国内外分离株、LaSota、F48E9、V4等相比，具有明显的地域性和种属性。 相似文献
5.
一株野禽新城疫强毒分离株的分子特性及其对不同宿主的致病性 总被引：1，自引：0，他引：1
新城疫(newcastle disease,ND)是禽类的两大疫病之一,新城疫病毒(Newcastle disease virus,NDV)是该病的病原体。为了解野禽NDV的分子特性、致病性及其与家禽NDV之间的关系,2011年分离到一株野禽NDV,将其命名为NDV2;通过标准生物学毒力检测证实其为强毒株。参照GenBank发布的NDV全长基因序列设计9对特异性引物,对NDV2进行全基因的序列测定和分析。拼接后序列分析表明,NDV2株的全基因组序列总长为15 192 nt,包含6个开放阅读框,分别编码6种蛋白nucleocapsid protein(NP)、phosphoprotein(P)、matrix protein(M)、fusion protein(F)、haemagglutinin-neuraminidase(HN)和large protein(L)的基因,长度分别为1 753、1 451、1 241、1 792、2 004和6 703 nt(GenBank登录号:KF306265)。与国内多数禽源分离毒株相似,NDV2毒株亦属于ClassⅡ、基因Ⅶ型。综合各个蛋白的同源性比较,NDV2株与近年国内流行的一些基因Ⅶ型毒株如:Egret.GX.11、Duck.WF.00、Goose.GD450.11等的同源性较高,远高于其他地区和其他基因型的毒株,这说明NDV2毒株与国内的野毒株亲缘性较高;与NDV2同源性差距最大的为ClassⅠ的JS10毒株。F基因的裂解位点显示:全基因长度为15 192nt的毒株(包括NDV2毒株在内)在裂解位点处的氨基酸序列为RRQKRF或KRQKRF,为强毒株序列。从基因分型结果来看,NDV2毒株和绝大多数参考毒株F基因和HN基因分型结果是一致的;唯一不同的就是印度鸡源分离株NDV4毒株,F基因分型它属于Ⅱ型,HN基因分型它属于Ⅶ型。分析各个基因的起始序列和终止序列后发现:NDV2株与其它参考NDV毒株的各个基因的起始序列完全相同,说明各基因起始序列保守性较高;不同之处是NDV2毒株F和HN基因的终止序列与La Sota和Sweden95一致,与其他野禽毒株不同。以NDV2人工感染无特定病原(Spicific pathogenfree,SPF)鸡(Gallus gallus)、鸭(Anas platyrhynchos platyrhynchos Linnaeus.)和鸽(Rupestris Pallas),并且每个攻毒组设同居感染组;来观察NDV2毒株对不同宿主的致病性。结果表明,NDV2毒株均可引起3种攻毒动物发病,剖解后肠道、腺胃、气管等处有淤血、出血现象;而且病毒对鸡和鸽的致病性明显高于鸭。同时NDV2毒株可引起50%同居鸡和20%同居鸽感染发病,不能引起同居鸭发病。本研究为NDV的遗传变异特征和致病规律等方面提供基础资料。 相似文献
6.
由于Bt Cry1Aa、Cry1Ab和Cry1Ac晶体蛋白之间具有很高的同源性(82%~90%),采用常规的单抗制备方法很难制取特异性强的Bt Cry1Ab单抗,为了制备抗Bt Cry1Ab蛋白的特异性单克隆抗体(MAB),本研究从NCBI获得了Bt Cry1Ab蛋白的氨基酸序列,根据ANTHEPORT和DNAStar软件对其抗原性、亲水性和表位性分析结果选定BtCry1Ab特异性肽段进行人工合成,并将其偶联于匙孔血蓝蛋白(KLH)免疫动物,应用细胞融合技术制备了抗该肽段的杂交瘤细胞22株。通过ELISA试验从中筛选出与Bt Cry1Ab天然蛋白产生特异性反应的单克隆抗体杂交瘤细胞株一株(3A10)。经检测,其分泌的抗体亚类为IgG1型;轻链属κ型;杂交瘤细胞株染色体数目为89~108条;用其制作的腹水对Bt Cry1Ab合成肽的反应效价为1∶1×107;对Bt Cry1Ab天然蛋白的反应效价为1∶1×104;纯化后的抗体对Bt Cry1Ab合成肽的效价为1∶1×108;对Bt Cry1Ab天然蛋白的效价为1∶2×104。抗体的相对亲和力为0.5μg/mL,对Bt Cry1Ab蛋白的最低可检测值为10ng/mL。ELISA结果显示,3A10杂交瘤细胞株所分泌的MAB能特异性识别合成肽和Bt Cry1Ab蛋白,而对同源的Cry1Ac和Cry1Aa蛋白无交叉反应;本研究所制备的Bt Cry1Ab单克隆抗体能够对常规棉和抗虫棉(Gossypium hirsutumL.)进行有效的区分,并且能特异性的识别其中的Bt Cry1Ab蛋白。 相似文献
7.
从噬菌体随机七肽库中筛选得到赭曲霉毒素A的模拟表位,并以其替代毒素标品建立赭曲霉毒素A免疫学快速检测方法。以纯化的抗赭曲霉毒素A单克隆抗体(OTA mAb)为配基,亲和筛选融合表达在丝状M13噬菌体次要衣壳蛋白(PⅢ)上的随机七肽库,以ELISA方法鉴定阳性克隆,并以筛选的模拟表位建立赭曲霉毒素A的ELISA检测方法。经过4轮亲和筛选,共得到22株与OTA mAb特异结合的阳性噬菌体克隆,且该结合都能够被OTA标品阻断,DNA测序及多肽核心序列分析后,得到赭曲霉毒素A的模拟表位的主要序列为:MPLWXDL,X为任意氨基酸,以阳性噬菌体克隆建立的竞争ELISA检测方法,线性范围为250～8000pg/ml。基于制备的OTA mAb,利用噬菌体随机七肽库可成功筛选到赭曲霉毒素A的模拟表位,并能够替代OTA标品建立直接竞争模式的免疫学快速检测方法。 相似文献
8.
人胰腺激肽释放酶cDNA的克隆、序列分析和原核表达 总被引：3，自引：0，他引：3
摘要: 为了研制人激肽释放酶（KLK1）特异单克隆抗体和进行重组酶的鉴定及纯化，根据已发表的人KLK1序列设计引物，用PCR扩增法从人胰腺单链cDNA库中特异地扩增出KLK1 cDNA。将其克隆入pGEM-T载体中,对5个重组质粒进行了序列测定，其中1个cDNA克隆的核苷酸序列与已发表的人肾/胰/唾液腺KLK1 cDNAs序列完全相同或有3个核苷酸差异。将去除信号肽序列的KLK1 cDNA以正确阅读框插入表达载体pGEX-4T-3,构建成重组质粒pGEX-KLK1。此重组质粒转化的E.coli 经IPTG诱导后表达分子量为48 kDGST-KLK1融合蛋白，表达产物主要以不溶性包涵体形式存在，可溶性部分能被Glutathione Sepharose 4B特异吸附，两者都能被GST特异单克隆抗体识别。用SDS-PAGE分离纯化的融合蛋白免疫小鼠，获得的抗血清的ELISA效价为1∶1600。结果表明，克隆的人KLK1 cDNA及其表达产物是正确的，可以用于人KLK1单克隆抗体的制备。 相似文献
9.
通过反转录-聚合酶链反应(RT-PCR)扩增了新城疫病毒(NDV)F48E8株核衣壳蛋白(NP)基因,并克隆到pUC18中,得阳性克隆pUCNP.应用分子克隆技术,将NP基因导入鸡痘病毒(FPV)转移载体pFG1175-1中P7.5启动子的下游,得到含NDV NP基因的质粒pFGNP1175-1.利用脂质体转染技术,将该质粒转染FPV疫苗株282E4感染3～4 h的鸡胚成纤维细胞,采用蓝斑筛选方法纯化多次,得到稳定的重组FPV.用狄高辛标记的探针进行斑点杂交实验,结果表明pFGNP1175-1已与FPV 发生同源重组;用抗NDV的SPF鸡的阳性血清进行间接免疫荧光实验,证实重组FPV在感染细胞中表达了F48E8株NP蛋白.该基因两端部分序列与已发表的D26株NP基因对应区域的核苷酸同源性为89.1%,推断的氨基酸同源性则为93.1%. 相似文献
10.
犬细小病毒病是危害养犬业的重要传染病之一,患病犬难以治愈。单克隆抗体治疗此病效果明显,本文介绍了制备抗CPV-2a单克隆抗体的方法。用纯化的犬细小病毒（canine parvovirus,CPV）2a型分离株免疫新西兰大白兔和Balb/c小鼠制备抗CPV-2a多克隆抗体及单克隆抗体。经亚克隆得到1H9、2B5、2B7和2C7共4株单抗,Western blotting鉴定单抗的免疫反应性;间接ELISA方法检测单抗的特异性。为了快速对犬细小病毒病作出诊断,建立了CPV-2a双抗夹心ELISA方法。兔多抗作为捕获抗体,鼠单抗作为示踪抗体,辣根过氧化物酶标记羊抗鼠IgG作为检测系统;捕获抗体和示踪抗体最佳稀释度分别为1：800和1：2000;检测系统最佳稀释度为1：4000。结果表明：所得4株单抗与pET-32a-VP2蛋白发生特异性反应,且与狂犬病毒（RV）、犬温热病毒（CDV）不交叉反应;建立的双抗夹心ELISA方法对病毒的最低检出量为4.375μg/mL,与美国RB试剂盒相比,符合率为95%。单抗制备为犬细小病毒病的治疗奠定了基础;双抗夹心ELISA方法的建立为疑似粪便样本提供了简单、快速和可靠的检测手段。 相似文献
11.
Kevin H. Sutton Lyall D. Simmons Jill B. Cummack Sarah J. Roberts 《Cereal Chemistry》2016,93(4):352-356
Epitopes on the α‐gliadins are known to give rise to immune responses that may lead to the development of celiac disease in genetically predisposed individuals. The reduction of epitope levels in wheat‐based products would likely benefit this group of consumers and also consumers with non‐celiac gluten sensitivity. Conventional breeding of wheats with lowered epitope levels will take time, but in this study we show for the first time that milling technology can be used to produce flour mill streams that are depleted in α‐20 gliadin epitopes. Fifteen mill streams from two New Zealand wheat cultivars, Sapphire (a biscuit wheat) and Monad (a bread wheat), were tested with reversed‐phase HPLC and an α‐20 gliadin epitope ELISA kit. The level of α‐20 epitope measured in Sapphire gliadins was significantly less than that found in Monad gliadins, even taking into account differences in total protein content. For both cultivars, compared with the straight‐run flour, the break flours had similar or significantly higher proportions of α‐20 epitope per unit of protein, whereas most of the reduction streams had significantly lower proportions of α‐20 epitope per unit of protein. Theoretically, combining selected (mainly reduction) flour streams may produce flour with ∼75% of the epitope content of the straight‐run flour. 相似文献
12.
Xiaomin Guo Junxian Guo Xiuquan Li Xinming Yang Lihui Li 《Genetic Resources and Crop Evolution》2010,57(8):1217-1225
The high molecular weight glutenin subunits (HMW-GS) can be used for wheat quality improvement. Two novel alleles (designated
1Dx1.5* and 1Dy12.2*, respectively) at the Glu-D1 locus were identified in the Chinese wheat landrace variety ‘Jiuquanjinbaoyin’ by comparison of subunit mobility with that
previously identified in several standard hexaploid wheats. The 1Dx1.5* and 1Dy12.2* genes were isolated using the allele-specific PCR primers and the complete open reading frames (ORFs) were obtained. Allele
1Dx1.5* consists of 2487 bp encoding a mature protein of 827 amino acid residues, whereas 1Dy12.2* consists 1980 bp encoding 658 residues. Comparisons of amino acid sequences analysis showed that 1Dx1.5* had higher similarity
with the HMW-GS isolated from the wheat related species (Aegilops tauchi Coss.) than from the bread wheat varieties (Triticum aestivum L.). The 1Dy12.2* amino acid sequence showed a generally similar to the 1Dy12* isolated from Chinese endemic wheats. Meanwhile,
the dough properties of the lines expressing (null, 7+8, 1.5*+10), (null, 7+8, 2+12.2*) and (null, 7+8, 2+12), respectively,
were measured by a Mixograph, which demonstrates that the alleles 1.5*+10 can be considered as having positive impact on dough
strength when compared with the alleles 2+12. In addition, the subunits 2+12.2* also showed a greater impact on dough strength
than 2+12. 相似文献
13.
Buckwheat is generally regarded as a nutritionally rich food source. However, earlier studies prove that it also causes allergies to subjects. Allergenic proteins with a strong IgE-binding activity have been identified in common buckwheat (CB) and a 24 kDa allergen (rTBa) in tartary buckwheat (TB). The objective of this research was to clone and express a novel allergen in tartary buckwheat and to evaluate its structure and immunological activity. The 1773 bp full-length cDNA was amplified and cloned from the total RNA of TB by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methods. Its nucleotide sequence had high similarity with legume-like 13S storage protein mRNA in CB. The deduced amino acid sequence included a putative signal peptide and 18 fragments as its epitope sites. The predicted full-length TB allergen sequence was found to have two domains, and the recombinant protein reacted with sera from patients with positive IgE binding to buckwheat and had a lower binding ability than the recombinant TBa and recombinant TBb (C- and N-terminal amino acid sequence of TBt codes for protein). This fact suggests that full-length TB allergen may hydrolyze to two domains in vivo, decreasing the IgE-binding ability. 相似文献
14.
Ivanciuc O Mathura V Midoro-Horiuti T Braun W Goldblum RM Schein CH 《Journal of agricultural and food chemistry》2003,51(16):4830-4837
The high incidence of food allergies, including oral allergy syndrome, represent major considerations when introducing new crops and foods. A new structural database of allergenic proteins, SDAP-Food, http://fermi.utmb.edu/SDAP/, has been developed to aid in predicting the IgE-binding potential of novel food proteins and cross-reactivities among known allergens. The site is designed to facilitate the first steps of a decision tree approach to determine the allergenicity of a given protein, based on the sequence and structural similarity to known allergens and their IgE binding sites. Immunological tests can then be used to confirm the predictions. A hierarchical procedure for identifying potential allergens, using a physical property-based sequence similarity index, has been designed to identify regions that resemble known IgE binding sites. As an example, SDAP tools were used to find food allergen sequences similar to an IgE binding site of the Jun a 3 allergen from mountain cedar pollen. The SDAP sequence similarity search matched the Jun a 3 epitope to regions in several food allergens, including cherry (Pru av 2), apple (Mal d 2) and pepper (Cap a 1), which are, like Jun a 3, members of the plant pathogenesis-related (PR-5) protein family. Homology modeling, using our EXDIS/DIAMOD/FANTOM program suite, indicated a similar surface location and structure for the potential epitope region on all of these allergens. The quantitative approach presented here can be used as part of a screening process for potential allergenicity of recombinant food products. 相似文献
15.
以纯化的原核表达的鸡贫血病毒结构蛋白为抗原，免疫6-8w的雌性Balb/c鼠, 经过三次免疫后，取其脾细胞与SP2/0骨髓瘤细胞进行融合，以纯化的蛋白为抗原进行ELISA检测，将阳性细胞孔经过三次有限稀释，共获得6株能稳定分泌特异性抗体的阳性细胞株。间接免疫荧光试验证实6株单抗可以与鸡贫血病毒感染的MDCC-MSB1细胞发生特异性反应，Western blot结果显示单抗与杆状病毒表达的VP1重组蛋白可以发生特异性反应。利用单克隆抗体亚型鉴定试剂盒进行亚型鉴定，结果表明6株单抗均为IgG1亚型，且所有单抗的轻链均为κ链。用单克隆抗体对分段表达的VP1蛋白进行免疫印迹分析，初步鉴定了筛选出的单抗所对应的抗原表位，其中有四株单抗识别的表位位于VP1 218-274位氨基酸之间，另外两株单抗识别的表位分别位于275-301位、324-369位氨基酸之间。 相似文献
16.
抗对硫磷基因工程四价抗体在大肠杆菌中高效表达与鉴定 总被引：1，自引：0，他引：1
为提高抗对硫磷基因工程四价抗体在大肠杆菌中可溶性表达量,本研究利用克隆技术得到四价抗体基因sc-sa,将该融合基因连接至表达载体pTO-T7的Ω序列和T7启动子下游,构建成功的表达质粒导入大肠杆菌OrigamiB(DE3),经IPTG诱导表达,SDS-PAGE和Western Blot鉴定表达产物,Ni亲和层析纯化蛋白,间接非竞争ELISA法测定该四价抗体的亲和力。结果表明在大肠杆菌OrigamiB(DE3)中表达分子量约为46kDa的四价抗体,0.1mmol/L的IPTG在30℃条件下诱导原核表达,外源蛋白占菌体总蛋白含量近50%,纯化后可溶性蛋白纯度在90%以上,ELISA结果显示该抗体与对硫磷结合呈阳性,抗体效价在1∶1×106以上,亲和常数为6.84×108L/mol。与母源抗体和相应单链抗体相比,利用pTO-T7载体四价抗体在大肠杆菌OrigamiB(DE3) 中可实现高效表达,且抗体效价和亲和力均得到进一步提高。 相似文献
17.
Although many sequences and linear IgE epitopes of allergenic proteins have been identified and archived in databases, structural and physicochemical discriminators that define their specific properties are lacking. Current bioinformatics tools for predicting the potential allergenicity of a novel protein use methods that were not designed to compare peptides. Novel tools to determine the quantitative sequence and three-dimensional (3D) relationships between IgE epitopes of major allergens from peanut and other foods have been implemented in the Structural Database of Allergenic Proteins (SDAP; http://fermi.utmb.edu/SDAP/). These peptide comparison tools are based on five-dimensional physicochemical property (PCP) vectors. Sequences from SDAP proteins similar in their physicochemical properties to known epitopes of Ara h 1 and Ara h 2 were identified by calculating property distance (PD) values. A 3D model of Ara h 1 was generated to visualize the 3D structure and surface exposure of the epitope regions and peptides with a low PD value to them. Many sequences similar to the known epitopes were identified in related nut allergens, and others were within the sequences of Ara h 1 and Ara h 2. Some of the sequences with low PD values correspond to other known epitopes. Regions with low PD values to one another in Ara h 1 had similar predicted structure, on opposite sides of the internal dimer axis. The PD scale detected epitope pairs that are similar in structure and/or reactivity with patient IgE. The high immunogenicity and IgE reactivity of peanut allergen proteins might be due to the proteins' arrays of similar antigenic regions on opposite sides of a single protein structure. 相似文献
18.
Martos G Pineda-Vadillo C Miralles B Alonso-Lebrero E López-Fandiño R Molina E Belloque J 《Journal of agricultural and food chemistry》2012,60(20):5215-5220
Riboflavin binding protein (RfBP) is a minor protein in hen egg; its potential involvement in egg allergy has seldom been studied. The aim of this work was to investigate the IgE binding capacity of RfBP before and after simulated gastrointestinal digestion. It was shown that digestion of RfBP mainly occurred during the gastric phase. The protein fragments resulting from the subsequent duodenal phase remained linked through disulfide bonds. Both the intact protein and its digests were subjected to inhibition ELISA with sera obtained from patients allergic to egg. The results revealed significant IgE binding to intact RfBP, whereas the digests showed reduced but substantial IgE binding levels, with serum-to-serum variability. The RfBP digests were then subjected to immunoblot with allergic patients' sera, and the IgE-reactive peptides were further analyzed by MALDI-TOF/TOF mass spectrometry for sequence determination. The RfBP sequence 41-84 was identified as a novel IgE binding peptide in patients allergic to egg. 相似文献
19.
运用RT-PCR技术克隆了水稻南方黑条矮缩病毒（southern rice black-streaked dwarf virus,SRBSDV）湖南鼎城株系的基因组S10片段（SRBSDV-HuNDCS10）,并对其全序列进行了测定和生物信息学分析。结果显示,SRBSDV-HuNDC S10片段全长为1797bp（登录号：JQ337964）,含有1个ORF,编码557个氨基酸残基的衣壳蛋白,推测分子量约62.6kD,推测等电点为7.62,与已报道的广东、海南和云南分离物病毒的S10作比较,它们的核苷酸相似性分别为99.7%、99.0%和98.4%,氨基酸相似性分别为100.0%、99.5%和99.3%。对SRBSDV-HuNDCS10及部分Fijiviruses病毒对应片段在5＇URT与3＇URT存在的保守序列和互补序列进行了归纳,对其ORF编码的氨基酸序列进行了motif查找,得到该属（Fijiviruses）氨基酸序列的10个保守区段。此外,进行了糖基化位点、磷酸化位点及B细胞抗原表位预测,发现了3个可能的N端豆蔻酰基化位点,可能与病毒的侵染机制有关。 相似文献
20.
Reduced immunogenicity of beta-lactoglobulin by conjugation with carboxymethyl dextran differing in molecular weight 总被引：2，自引：0，他引：2
Kobayashi K Hirano A Ohta A Yoshida T Takahashi K Hattori M 《Journal of agricultural and food chemistry》2001,49(2):823-831
To reduce the immunogenicity of beta-lactoglobulin (beta-LG), two beta-LG-carboxymethyl dextran (CMD) conjugates (Conj. 40 and Conj. 162) were prepared by using water-soluble carbodiimide (EDC). The molar ratios of beta-LG to CMD in Conj. 40 and Conj. 162 were 8:1 and 7:1, respectively. Each conjugate maintained approximately 50% of the retinol binding activity of beta-LG. Structural analyses by intrinsic fluorescence, CD spectra, and ELISA with monoclonal antibodies indicated that the surface of beta-LG in each conjugate was covered by CMD without great disruption of native conformation. By conjugation with CMD, the antibody response to beta-LG was reduced in BALB/c, C3H/He, and C57BL/6 mice, which was eminent in Conj. 162. The results of B cell epitope scanning using overlapping synthesized peptides showed that the linear epitope profiles of the conjugates were similar to those of beta-LG, whereas the antibody response to each epitope was reduced, which was eminent in Conj. 162. It was concluded that conjugation with CMD of higher molecular weight is effective in reducing the immunogenicity of beta-LG and that masking of epitopes by CMD is responsible for the reduced immunogenicity. 相似文献