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1.
Protection of mice against the Lyme disease agent by immunizing with recombinant OspA 总被引：119，自引：0，他引：119
Lyme borreliosis is a tick-borne illness caused by Borrelia burgdorferi. The gene for outer surface protein A (OspA) from B. burgdorferi strain N40 was cloned into an expression vector and expressed in Escherichia coli. C3H/HeJ mice actively immunized with live transformed E. coli or purified recombinant OspA protein produced antibodies to OspA and were protected from challenge with several strains of B. burgdorferi. Recombinant OspA is a candidate for a vaccine for Lyme borreliosis. 相似文献
2.
A fundamental tenet of microbial pathogenesis is that bacterial pathogens must overcome host iron limitation to establish a successful infection. Surprisingly, the Lyme disease pathogen Borrelia burgdorferi has bypassed this host defense by eliminating the need for iron. B. burgdorferi grew normally and did not alter gene expression in the presence of iron chelators. Furthermore, typical bacterial iron-containing proteins were not detected in cell lysates, nor were the genes encoding such proteins identified in the genome sequence. The intracellular concentration of iron in B. burgdorferi was estimated to be less than 10 atoms per cell, well below a physiologically relevant concentration. 相似文献
3.
莱姆病(Lyme disease,LD)是一种由蜱传播的人兽共患病,对人类健康和畜牧业的发展有严重危害,已经被世界卫生组织列为应重点防治研究的传染病。黑龙江省是我国最先发现本病的地区,在本地区进行蜱现场防制试验、动物及圈舍蜱防制试验、蜱驱避试验。结果表明,在蜱密度较高的林区,用0.5%毒死蜱和0.045%高效氯氰菊酯喷洒,可将蜱杀灭,持效在30 d以上;用同样药物对动物及其圈舍喷洒,24h可杀灭所有的蜱;避蚊胺作为林场作业工人和旅行者个人防护用药具有较好效果。 相似文献
4.
The genetics of spirochetes, a division of eubacteria, has been little studied. Double-stranded linear plasmids were found in Borrelia burgdorferi, the agent of Lyme disease. A 49-kilobase linear plasmid contained the ospA and ospB genes, which encode the major outer membrane proteins of strain B31. Molecules of the 49-kilobase plasmid rapidly reannealed after alkaline denaturation; rapid renaturation was prevented if the 49-kilobase plasmids were first treated with S1 nuclease. When denatured plasmid molecules were examined directly, single-stranded circles of approximately 100-kilobase circumference were seen. These studies provide direct visual evidence that the linear plasmids have covalently closed ends. This form of DNA occurs in some animal viruses, but it has not heretofore been described in prokaryotic organisms. 相似文献
5.
A single recombinant plasmid expressing two major outer surface proteins of the Lyme disease spirochete 总被引：48，自引：0，他引：48
A gene bank of DNA from the Lyme disease spirochete was constructed in the plasmid pBR322. Plasmid pTRH32, a recombinant that in Escherichia coli expresses the two major outer surface proteins of the Lyme disease spirochete, was identified. One of the recombinant products, designated OspA, represents a surface protein that appears to be common to all Lyme disease spirochetes, whereas the other recombinant product, designated OspB, represents a more variable surface protein. This recombinant plasmid provides a foundation for future studies on the epidemiology and pathogenesis of Lyme disease as well as on the genetic organization of the etiologic agent. 相似文献
6.
DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells 总被引：140，自引：0，他引：140
C Y Ou S Kwok S W Mitchell D H Mack J J Sninsky J W Krebs P Feorino D Warfield G Schochetman 《Science (New York, N.Y.)》1988,239(4837):295-297
By means of a selective DNA amplification technique called polymerase chain reaction, proviral sequences of the human immunodeficiency virus (HIV-1) were identified directly in DNA isolated from peripheral blood mononuclear cells (PBMCs) of persons seropositive but not in DNA isolated from PBMCs of persons seronegative for the virus. Primer pairs from multiple regions of the HIV-1 genome were used to achieve maximum sensitivity of provirus detection. HIV-1 sequences were detected in 100% of DNA specimens from seropositive, homosexual men from whom the virus was isolated by coculture, but in none of the DNA specimens from a control group of seronegative, virus culture-negative persons. However, HIV-1 sequences were detected in 64% of DNA specimens from seropositive, virus culture-negative homosexual men. This method of DNA amplification made it possible to obtain results within 3 days, whereas virus isolation takes up to 3 to 4 weeks. The method may therefore be used to complement or replace virus isolation as a routine means of determining HIV-1 infection. 相似文献
7.
从福尔马林保存的牙鲆中克隆的18S rDNA进行系统发育分析的可靠性 总被引：1，自引：0，他引：1
从福尔马林保存的牙鲆和活体牙鲆肌肉组织中，利用基因拼接的方法首次克隆到1205bp和1295bp的长片段序列。福尔马林保存标本与活体标本DNA序列的同源率为82％；和活体标本的18S rDNA序列相比，标本DNA的碱基缺失率为7％，碱基替换比例为11％。利用福尔马林标本的18S rDNA同源性高的区域进行系统发育分析，表明其结果是可信的。 相似文献
8.
从福尔马林保存的牙鲆和活体牙鲆肌肉组织中，利用基因拼接的方法首次克隆到1205bp和1295bp的长片段序列。福尔马林保存标本与活体标本DNA序列的同源率为82％；和活体标本的18S rDNA序列相比，标本DNA的碱基缺失率为7％，碱基替换比例为11％。利用福尔马林标本的18S rDNA同源性高的区域进行系统发育分析，表明其结果是可信的。 相似文献
9.
V Manzari A Gismondi G Barillari S Morrone A Modesti L Albonici L De Marchis V Fazio A Gradilone M Zani 《Science (New York, N.Y.)》1987,238(4833):1581-1583
A new human retrovirus was isolated from a continuous cell line derived from a patient with CD4+ Tac- cutaneous T cell lymphoma/leukemia. This virus is related to but distinct from human T cell leukemia/lymphoma virus types I and II (HTLV-I and HTLV-II) and human immunodeficiency virus (HIV-1). With the use of a fragment of provirus cloned from one patient with T cell leukemia, closely related sequences were found in DNA of the cell line and of tumor cells from seven other patients with the same disease; these sequences were only distantly related to HTLV-I. The phenotype of the cells and the clinical course of the disease were clearly distinguishable from leukemia associated with HTLV-I. All patients and the wife of one patient showed a weak serological cross-reactivity with both HTLV-I and HIV-1 antigens. None of the patients proved to be at any apparent risk for HIV-1 infection. The name proposed for this virus is HTLV-V, and the date indicate that it may be a primary etiological factor in the major group of cutaneous T cell lymphomas/leukemias, including the sporadic lymphomas known as mycoses fungoides. 相似文献
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11.
DNA sequences mediating class switching in alpha-immunoglobulins 总被引：49，自引：0，他引：49
Immunoglobulin class switching involves specific DNA rearrangements of the gene segments coding for heavy chain constant regions (CH) during B lymphocyte differentiation. In two different cases of C mu to C alpha switching examined here (T15 and M603) and one taken from the literature (MC101), three different sites on the 5' side of C mu and three different sites on the 5' side of C alpha are joined together in the process of CH switching. The sequences surrounding the three germ-line C alpha sites of recombination are highly conserved blocks of 30 nucleotides that may serve as recognition sequences for CH switching to the C alpha gene. This putative recognition sequence is repeated 17 times in approximately 1400 nucleotides of the germ-line Calpha 5' flanking sequence. The lack of homology between this C alpha sequence and sequences reported for the C gamma 1 and C gamma 2b switch sites suggests that heavy chain switching is mediated by class-specific recognition sequences and, presumably, class-specific regulatory mechanisms. In addition, it appears that in one example (MC101) CH switching progressed from C mu to C alpha to C gamma 1. This switching pathway may present difficulties for the simple deletional model of CH switching. 相似文献
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13.
从猪血中分离附红细胞体,提取其基因组DNA,用EcoRI对基因组DNA进行酶切,将酶切片段进行琼脂糖电泳,得到一个1.8 kb的DNA片段,根据DNA序列分析与GenBank已知基因比较和PCR方法确定该片段具有特异性,以该片段为模板进行PCR,并将其扩增产物与其它猪的其他细菌DNA扩增产物相比较,建立一种检测猪外周血中附红细胞体的特异性PCR检测方法,并通过斑点杂交试验检测该产物特异性,通过临床应用检验该法的适用性。结果表明:扩增产物大小为782 bp,其序列与已知附红细胞体序列同源性为100%,斑点杂交试验证实该产物为附红细胞体特异性产物,将该方法应用于检测10头临床症状典型的感染猪和10头健康猪的血液,所有感染和隐性带菌猪的PCR检测结果为阳性,健康猪为阴性。猪外周血中附红细胞体的特异性PCR检测方法适用于检测无临床症状的隐性带菌动物。 相似文献
14.
Isolation of infectious bovine rhinotracheitis virus from the soft-shelled tick, Ornithodoros coriaceus 总被引：1，自引：0，他引：1
Infectious bovine rhinotracheitis virus was isolated from the soft-shelled tick (Ornithodoros coriaceus). Serological identification of the isolated viruses was confirmed by restriction endonuclease digestion of purified virus DNA. These isolations indicate that the soft-shelled tick may be a vector for infectious bovine rhinotracheitis virus. This may be the first reported isolation of mammalian herpesvirus from an arthropod vector. 相似文献
15.
MT Maurano R Humbert E Rynes RE Thurman E Haugen H Wang AP Reynolds R Sandstrom H Qu J Brody A Shafer F Neri K Lee T Kutyavin S Stehling-Sun AK Johnson TK Canfield E Giste M Diegel D Bates RS Hansen S Neph PJ Sabo S Heimfeld A Raubitschek S Ziegler C Cotsapas N Sotoodehnia I Glass SR Sunyaev R Kaul JA Stamatoyannopoulos 《Science (New York, N.Y.)》2012,337(6099):1190-1195
16.
依据Gen Bank中注册的鸡马立克氏病病毒(MDV)、鸡传染性喉气管炎病毒(ILTV)、鸡痘病毒(FPV)基因组核苷酸序列中的保守序列设计引物,3种病毒各设计3对引物,共计9对引物,经生物学软件筛选后,得到3对相互匹配较佳的引物。3对匹配引物经单一PCR验证后,优化复合PCR反应条件,确定最佳的退火温度、引物浓度和Taq DNA聚合酶浓度;经特异性、敏感性试验及简化PCR试验,建立简易复合PCR检测方法。复合PCR扩增出的3条带大小与预期一致,即228 bp(MDV)、400 bp(ILTV)、499 bp(FPV);建立的复合PCR方法能同时检测出100 fg MDV、ILTV、FPV的DNA。同时依据PCR技术原理,将经典的三温热循环改进为二温热循环试验,即将退火和延伸合并为一步(62℃),缩短了反应时间。建立的3种病毒的复合PCR检测方法具有较高的特异性和敏感性,可以用于临床病料的快速诊断。 相似文献
17.
二温式PCR检测猪传染性胸膜肺炎放线杆菌方法的建立与应用 总被引：9，自引：0，他引：9
为了探索简便、快速确诊猪传染性胸膜肺炎的方法,根据猪传染性胸膜肺炎放线杆菌(APP)apx IV基因序列,设计合成1对特异性引物,建立聚合酶链反应(PCR)检测APP的方法。采用该方法对APP和其他6种猪病病原核酸进行检测,结果只对APP扩增出与预期大小相符的422bp DNA片段,而对其他6种猪病病原核酸的扩增结果为阴性。该PCR最低可检出10pg的APP DNA。对送检的46份可疑病猪组织进行检测,结果有19份样品为阳性,27份为其他病原感染。 相似文献
18.
Scissors-grip model for DNA recognition by a family of leucine zipper proteins 总被引：152，自引：0，他引：152
C/EBP is a sequence-specific DNA binding protein that regulates gene expression in certain mammalian cells. The region of the C/EBP polypeptide required for specific recognition of DNA is related in amino acid sequence to other regulatory proteins, including the Fos and Jun transforming proteins. It has been proposed that these proteins bind DNA via a bipartite structural motif, consisting of a dimerization interface termed the "leucine zipper" and a DNA contact surface termed the "basic region." An evaluation of the properties of conserved amino acids within the basic region of 11 deduced protein sequences, coupled with the observation that they are located at an invariant distance from the leucine zipper, has led to the formulation of a "scissors-grip" model for DNA binding. The architectural features of this model are well suited for interaction with directly abutted, dyadsymmetric DNA sequences. Data supportive of the model were obtained with chemical probes of protein: DNA complexes. 相似文献
19.
猪巨细胞病毒的PCR检测及其gB基因特征分析 总被引：1，自引：0，他引：1
根据GenBank登录的猪巨细胞病毒(PCMV)gB基因序列设计检测引物和全长gB基因引物,对来自湖南浏阳和江西景德镇不同养殖场的猪鼻拭子样品(51份)进行PCR检测,60.78%(31/51)的鼻拭子样品扩增出了260 bp的特异性条带.再以阳性样品为模板扩增全长gB基因,并将其克隆到pMD18-T载体,核苷酸序列测定和结果分析表明,所获得PCMVgB基因序列全长2 580 bp,编码860个氨基酸,与国外PCMV分离株的核苷酸同源性为97.6% ～ 98.9%,氨基酸同源性为97.0%～98.6%,且 PCMV可分为2个群:一群为中国湖南株、英国株和德国株,命名为A群;另一群为日本株、瑞士株和西班牙株,命名为B群.抗原表位优势、亲水性、表面可及性预测分析表明,PCMVgB蛋白是理想的免疫学抗原候选蛋白. 相似文献
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以谷子(Setaria italica Beauv.)抗锈病植株十里香和感锈病植株豫谷1号为材料,利用抗病基因同源序列(RGA)技术克隆谷子核苷酸结合位点(NBS)类型RGA并进行分析.共获得了3个RGA,分别命名为RUS1-1、RUS1-2、RUS1-3(Resistance against Uromyces setariae-italicae,RUS),它们编码的蛋白质均含有NBS类型抗病基因编码蛋白的共有特征结构域P-loop和Kinase2α.BLASTX分析结果表明,3个RGA的编码蛋白与水稻中含有NB-ARC信号传导结构域的蛋白质、NBS-LRR类型抗病基因等的编码蛋白同源性为47％～66％.聚类分析结果表明,3个RGA属于NBS-LRR类型抗病基因,且与番茄NBS-LRR类型抗病基因12聚为一类. 相似文献