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1.
于氟氰菊酯分子的酸部分连接4-氨基丁酸间隔臂(HCA1)、醇部分连接丁二酸酐间隔臂(HCB2),分别合成了两种不同的半抗原,通过碳二亚胺法将HCA1和HCB2分别与牛血清蛋白(BSA)偶联制备了人工抗原HCA1-BSA和HCB2-BSA,通过混合酸酐法将HCA1、HCAS 和HCB2分别与卵清蛋白(OVA)偶联制备包被抗原,两种人工抗原通过免疫新西兰大白兔制备相应的多克隆抗体。结果表明,用HCA1-BSA获得的多克隆抗体对氯氟氰菊酯和氯氟氰菊酸有免疫反应性(IC50值分别为33.12 和0.95 mg/L),用HCB2-BSA获得的多克隆抗体对氯氟氰菊酯几乎没有免疫反应性。 相似文献
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以除草剂2甲4氯(MCPA,简写为M)、氯化亚砜、氨基己酸和氨基丁酸等为起始原料,经两步化学反应分别合成了两种MCPA半抗原:6-(2-甲基-4-氯苯氧乙酰基)氨基己酸(MC)和4-(2-甲基-4-氯苯氧乙酰基)氨基丁酸(MB)。并通过碳二亚胺法和混合酸酐法将MCPA、MC和MB分别与载体蛋白(BSA、OVA)偶联制备了2甲4氯的免疫抗原和包被抗原。再将M-BSA、MC-BSA和MB-BSA分别免疫新西兰大白兔获得了3种抗2甲4氯的多克隆抗体:抗M-BSA抗体(M-Ab)、抗MC-BSA抗体(MC-Ab)和抗MB-BSA抗体(MB-Ab),当相应的包被抗原浓度均为4 μg/mL时,相应的冻干粉效价分别为1.0×106,1.2×106和2.0×106。在同源反应中,仅M-Ab适用于2甲4氯的残留检测,其线性浓度范围为0.01~10 mg/L(IC50为1.3 mg/L,MC-Ab 和MB-Ab均大于50 mg/L),最低检测限为0.01 mg/L。研究结果为进一步研制2甲4氯快速检测试剂盒奠定了基础。 相似文献
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通过乙草胺与3-巯基丙酸在碱性条件下反应合成了半抗原——乙草胺-巯基丙酸(AMPA)。采用活性酯法将半抗原AMPA分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联制备了人工抗原。通过紫外光谱测定了AMPA与BSA及OVA间的分子结合比分别为41和26。用AMPA-BSA免疫小鼠,制备得到的单克隆抗体的效价为1∶ 16×104。以AMPA-OVA作包被抗原,用乙草胺单克隆抗体建立了ELISA检测方法,IC50值为0.55 μg/L,方法的检测范围为0.04~5 μg/L,检测限为0.04 μg/L。 相似文献
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用 ELISA 法鉴定了小麦近缘种赖草属(Leymus)、披碱草属(Elymus)、鹅冠草属(Roegneria)3个属的21个种,其中17个种抗 BYDV。21145份小麦品种中筛选到症状轻、病毒含量高的耐病品种忻县冬麦、江西早等29份。3604份大麦品种中筛选到症状轻、病毒含量低的抗病品种C13208、小麦近缘种(Agropyronintemedium)和普通小麦杂交的异源八倍体中4无芒,中5,远中7,陇远45、46,远中1001,忻4079以及附加系 L1。现已获得抗 BYDV 的以中4无芒、L1为亲本的杂交后代。 相似文献
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为了研究和分离有机磷降解酶及其编码基因,从农药厂污水处理池的活性污泥中分离出一株可以高效降解甲基对硫磷的细菌 1-7, 经16S rDNA鉴定为假单胞菌Pseudomonas sp.。通过构建基因组文库的方法克隆了 1-7 的甲基对硫磷水解酶基因 ophc3, 该基因全长为975 bp,编码324个氨基酸,其中前24个氨基酸残基可能为信号肽序列。将其在大肠杆菌中表达,并对重组甲基对硫磷水解酶(OPHC3)进行纯化和酶学性质的研究结果表明,其酶促反应最适pH值为8.0,在pH 6.0~10.0的范围内放置30 min酶的相对活性均在70%以上;最适反应温度为45 ℃,但该酶不耐高温,60 ℃下保温10 min,相对活性降至46.77%。 相似文献
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Li Xianchun Wang Yinchang Zhang Qiansong Yu Ganjun Zhang Dunyang Yang Yantao Zhang Zhi Zhang Jianping Luo Shoushan Chen Caixia Ding Shiyin 《Pest management science》1997,50(3):183-186
Filter-paper residual toxicities of some insecticides used extensively in China were determined during 1994 using newly hatched (within 30 min) larvae of four Pectinophora gossypiella (Saunders) strains. The strains were field collections collected in the Yangtze River cotton-belt areas. Compared with the susceptible laboratory strain from Qunli (Lishui County, Jiangsu province), the four field strains from Anqing (Anhui province), Jiangling (Hubei province), Cixi(Zhejiang province) and Tongzhou(Jiangsu province) had developed 185-, 6·7-, 698- and 249-fold resistance, respectively, to deltamethrin. Cixi and Tongzhou field strains had also developed 103- and 94-fold resistance to fenvalerate, and 10- and 3·6- fold resistance to parathion-methyl. Percentage of survivors at diagnostic dosage for deltamethrin showed that the strains from Anqing, Jiangling, Cixi and Tongzhou had 87·2, 18·3, 90·1 and 74·6% resistant individuals respectively. Cixi and Tongzhou field strains had 88·9 and 65·3% resistant individuals after application of parathion-methyl, which was consistent with the corresponding resistance ratios. Studies of the effect of synergists piperonyl butoxide (PBO), triphenyl phosphate (TPP) with deltamethrin and parathion-methyl in Cixi, Anqing and Tongzhou field strains suggested that metabolic resistance mechanisms such as carboxylesterases (CarE) and mixed function oxygenases (MFO) were involved in parathion-methyl resistance, but not in deltamethrin resistance. © 1997 SCI. 相似文献
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Biotin conjugates are of considerable value in investigating the mode of action of biologically active compounds. Two biotin conjugates related to the antifungal compound cymoxanil [1-(2-cyano-2-methoximinoacetyl)-3-ethyl urea] were prepared as the first step in an effort to employ display cloning to identify the compound's target site. In the first conjugate, prepared in five steps, the biotin moiety was attached at the position occupied by the ethyl group in cymoxanil. In the second conjugate, prepared in four steps, the biotin moiety was attached through the oxime functional group. 相似文献
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几种危险性病毒抗血清的制备及检测方法的建立 总被引:1,自引:2,他引:1
番茄环斑病毒、烟草环斑病毒和南芥菜花叶病毒用改进的方法提纯后免疫兔子,制备出高质量的抗血清。琼脂扩散效价为1/320,用SPA-ELISA方法检测病汁液可达10-3~10-4。3种血清之间无交叉反应,与健康寄主和常见的自然寄主蛋白也无反应。应用3种血清分别对23种植物100多个采集的样品和进口种子进行了SPA-ELISA检测,未发现有上述3种病毒存在 相似文献
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为制备并鉴定番茄溃疡病菌(Clavibacter michiganensis subsp.michiganensis,Cmm)的单克隆抗体(McAbs),用全菌皮下免疫BALB/C小鼠,采用B细胞杂交瘤技术,经免疫、融合、间接ELISA筛选和克隆等,获得稳定分泌抗体的阳性杂交瘤细胞株,得到了抗番茄溃疡病菌的单克隆抗体。经免疫后获得3株单抗分别为1A4、1C3和1B7,经亚类鉴定分别是IgM、IgG1、IgG1;纯化腹水间接ELISA效价分别为1:3.2×106、1:8.1×105、1:3.2×106;与其他同属不同亚种无交叉反应。结果表明:3株单克隆抗体均具有较高特异性和敏感性,可作为番茄溃疡病菌的检测抗体,其中,1A4的效果最好。番茄溃疡病菌单克隆抗体的获得为进一步研发番茄溃疡病检测试剂盒奠定了基础。 相似文献
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Production of Monoclonal Antibodies against Apple Proliferation Phytoplasma and their Use in Serological Detection 总被引:2,自引:0,他引:2
Nazia Loi Paolo Ermacora Luigi Carraro Ruggero Osler Tseh An Chen 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):81-86
Two monoclonal antibodies were obtained against the apple proliferation phytoplasma that provide easy, rapid, specific and sensitive serological detection. They reacted specifically by using ELISA and immunofluorescence techniques with apple proliferation-infected periwinkles and apple trees from different regions in northern Italy and Slovenia, but not with several other phytoplasma isolates. We did not observe any monoclonal antibody reaction even using phytoplasmas belonging to the same phylogenetic group such as European stone fruit yellows and pear decline. Two serological techniques, immunofluorescence and ELISA, were compared with DAPI staining and PCR. From July until leaf fall ELISA was as sensitive as PCR but was more rapid and convenient than PCR; immunofluorescence was useful for specific detection of apple proliferation phytoplasma on roots throughout the year. Serological techniques could be conveniently applied in the roots, stems and leaves of apple trees depending on specific phenological stages of the plants. 相似文献
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The molecular basis of the genotoxicity of the commonly used organophosphorus insecticide, parathion-methyl is poorly understood and there is a lack of information on the possible effects of its metabolic conversion products. In the present work the action of parathion-methyl and its immediate metabolite paraoxon-methyl on DNA in human lymphocytes was compared using the comet assay. Parathion-methyl at 25 and 75 µM did not cause any significant changes but at 200 µM a significant increase in the tail moment was observed as compared with the control. Paraoxon-methyl at 25, 75 and 200 µM evoked dose-dependent DNA damage measured as a significant increase in comet tail moment of lymphocytes. The change evoked by paraoxon-methyl at 200 µM was much more pronounced than that by parathion-methyl at the same concentration. To search for the mechanism underlying the observed effect, the action of a well-known antioxidant, vitamin C, along with parathion-methyl and paraoxon-methyl was studied. The vitamin at 10 and 50 µM reduced the DNA-damaging activity of paraoxon-methyl at all its concentrations. The results indicate that the reported genotoxic effects of parathion-methyl could be mainly attributed to its metabolite paraoxon-methyl. The protective action of vitamin C suggests that paraoxon-methyl may cause oxidative DNA damage. © 1999 Society of Chemical Industry 相似文献
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I. Tóbiás D. Z. Maat H. Huttinga 《European journal of plant pathology / European Foundation for Plant Pathology》1982,88(5):171-183
Two Hungarian virus isolates from sweet pepper (K8) and melon (S4) were identified as cucumber mosaic virus (CMV) on the basis of host plant reactions and serology. The isolates were purified and antisera prepared. Homologous antiserum titers in double-diffusion tests were 256 (K8) and 512 (S4). They were serologically closely related to each other and to other CMV isolates. On the basis of symptoms they belong to different symptomatological groups of CMV; this was supported by serological properties. Sedimentation coefficients were c. 93 S, at 2 mg ml–1. Purified preparations, stained with 2% uranyl acetate, showed spherical particles. In ELISA purified preparations reacted with each other's antisera.Samenvatting Twee hongaarse virusisolaten uit paprika (K8) en meloen (S4) werden geïdentificeerd als komkommermozaïekvirus (CMV) met behulp van toetsplanten en serologie. Beide isolaten werden gezuiverd en er werden antisera tegen bereid. De homologe titers van de antisera in de agar-geldiffusietoets bedroegen 256 (K8) en 512 (S4). K8 en S4 waren serologisch nauw verwant aan elkaar, evenals aan andere CMV-isolaten. Op grond van hun symptomen op toetsplanten behoren ze tot verschillende symptomatologische groepen van CMV. Dit laatste werd gesteund door de serologische eigenschappen. Beide isolaten hebben een sedimentatiecoëfficiënt van ca 93 S, bij een concentratie van 2 mg ml–1. Gezuiverde preparaten, gekleurd met 2% uranylacetaat, bleken bolvormige deeltjes te bevatten. In ELISA reageerden gezuiverde preparaten van K8 en S4 met elkaars antisera. 相似文献
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甘肃省大丽花病毒病病原鉴定及病生理研究 总被引:1,自引:0,他引:1
经对受病毒感染的甘肃省临洮县大丽花进行间接ELISA和鉴别寄主测定,结果表明黄瓜花叶病毒(CMV)是其病原之一。田间调查表明大丽花病毒病在甘肃临洮县发生普遍,发病率达36%;其主要症状有花叶、卷叶花叶、蕨叶和矮缩。不同大丽花品种之间病毒病发病率有一定的差异,在调查的8个品种中‘鹦嘴红’发病率最低,只有7.43%,而‘洮阳荷花’发病率最高,达到42.47%。大丽花感染病毒后叶绿素(a+b)比正常的下降了23.61%;在叶片组织病变观察中,发现叶片感染病毒后叶绿体内的淀粉颗粒含量增多且肿大,油粒增多,几乎占满整个叶绿体结构。 相似文献
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从全国14个地区多种作物田采集土样56份,经室内分离、纯化及筛选,获得了对黄瓜褐斑病菌Corynespora cassiicola具有拮抗作用的16株细菌,其中拮抗作用最强的为菌株SDYT-79,其对黄瓜褐斑病菌的抑菌带宽度达37mm。室内盆栽试验结果显示,菌株SDYT-79对黄瓜褐斑病的防效达58.4%。孢子萌发试验表明,该菌株对黄瓜褐斑病菌孢子萌发有较强的抑制作用,其发酵无菌滤液及10倍稀释液对孢子萌发的抑制率均达100%。黄瓜褐斑病菌孢子在100倍、1000倍稀释无菌滤液中虽能萌发,但芽管形态异常。抑菌谱试验表明,该菌株对供试的11种植物病原真菌均有较好的抑菌作用,抑菌带在19.9~41.9mm之间。生理生化鉴定及16S rDNA序列测定结果表明该菌株为地衣芽孢杆菌Bacillus licheniformis。 相似文献