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《华北农学报》2015,(4)
为了鉴定类猪圆环病毒P1 VP1蛋白能否在体外自组装成病毒样颗粒(VLPs),首先PCR扩增P1 ORF1基因,利用p Fast BacTM1载体构建重组质粒,通过脂质体转染在Sf9细胞中表达P1 ORF1基因。然后通过RT-PCR技术及Western Blot分别从转录和蛋白质水平上鉴定该基因的表达,应用透射电镜观察该蛋白质是否形成了VLPs。结果显示,转染Sf9细胞后,第5天开始出现细胞病变;从接毒细胞中提取总RNA,经Recombinant DNaseⅠ处理除去基因组DNA后,RT-PCR能够扩增出362 bp的目的片段;收获接毒后72 h的细胞,Western Blot分析显示,P1 ORF1重组病毒样品能够与抗P1 VP1单抗发生特异性反应;电镜观察发现,该蛋白在Sf9细胞中形成了近椭圆形的包涵体。利用杆状病毒表达载体系统成功地在Sf9细胞中表达了类猪圆环病毒P1的ORF1基因,并证实在本试验条件下P1 VP1蛋白不能自组装成VLPs,为进一步研究P1 VP1蛋白的免疫学特性奠定基础。 相似文献
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类圆环病毒因子P1感染对猪外周血T淋巴细胞含量和增殖活性的影响 总被引:1,自引:0,他引:1
探讨类猪圆环病毒因子P1感染对猪细胞免疫功能的影响.将12头30日龄广西巴马小型猪随机分成试验组和对照组,每组6头.在猪感染P1后第3,8,17,24,31和40天,采用血常规和单色流式细胞术分析猪外周血T淋巴细胞的含量,通过WST法对其增殖活性进行检测.P1感染组的T淋巴细胞数量(绝对数和相对数)整体趋势低于对照组,而两组的T淋巴细胞增殖活性没有显著差别.P1感染对机体的细胞免疫功能有一定程度的影响. 相似文献
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为探讨类猪圆环病毒2型因子P1感染对猪抗病毒蛋白分子mRNA转录的影响,将P1分子克隆接种30日龄健康广西巴马小型猪,利用实时荧光定量PCR技术对猪外周血单个核细胞抗病毒蛋白分子PKR、2'-5'OAS2、 RNase L和Mx1的mRNA转录水平的变化进行了定量分析。结果表明,PKR mRNA转录在接种后8 d和40 d呈上调趋势;而2'-5'OAS2 mRNA在3 d和17 d下调,Mx1 mRNA也在接种后17 d下调;RNaseL mRNA转录水平则没有发生明显变化。研究揭示P1感染猪后,机体的抗病毒蛋白的功能受到了不同程度的影响。 相似文献
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【研究目的】为了研究猪圆环病毒的分子生物学特征及其发病机理和致病机制,【研究方法】将疑似猪圆环病毒2型的猪脾脏、淋巴结用胰蛋白酶消化处理后接种PK-15细胞,盲传6代后,病毒已经适应PK-15细胞,【研究结果】然后设计一对特异性引物利用PCR技术扩增出ORF2片段中540 bp的基因。第6代细胞病毒液经间接免疫荧光检测表明,在荧光显微镜下可见PCV特征的荧光斑,表明毒株具有感染性。【结论】将病毒纯化后进行电镜观察,病毒粒子直径约17nm,圆形,无囊膜,研究结果表明分离的病毒为猪圆环病毒。 相似文献
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类猪圆环病毒因子P1感染对外周血单个核细胞Toll样受体mRNA转录的影响 总被引:2,自引:0,他引:2
采用实时定量反转录多聚酶链反应方法,检测健康对照组及类圆环病毒因子P1感染组的猪外周血单个核细胞中TLRs mRNA的表达。结果表明,病毒感染后3 d,除TLR2和TLR10外,其余TLRs mRNA表达皆下调。此后不同时相感染组的TLRs mRNA表达水平基本都处于恢复、上调地位。具有明显变化的表现为感染后第24天和第40天,TLR2 mRNA的表达显著上调(P0.05);此外,感染后第31天和第40天的TLR9 mRNA表达也呈现上调趋势(P0.1)。结果提示,TLR2和TLR9介导的炎性反应可能在P1识别及其致病机制中起重要作用。 相似文献
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猪圆环病毒病是以断奶仔猪多系统衰竭综合征等为主要特征,并能引发多种疾病,对养猪业造成了严重威胁。
一、病原与流行特点
猪圆环病毒(PCV)在猪体内变异产生PCV—Ⅰ型和PCV—Ⅱ型,近年来流行的猪圆环病毒病被确定为PCV-Ⅱ型感染所致。PCV—Ⅱ型既可水平传播又可垂直传播。 相似文献
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Jens Jensen 《Euphytica》1979,28(1):47-56
Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans. 相似文献
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G. H. Kroon 《Euphytica》1994,76(1-2):125-125
Summary
K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes. 相似文献
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Hongyan WEI Jian ZHOU Jinguang LU Jingzheng SONG Qiongxi YU Zhihong JIANG 《Medicinal Plant》2019,(6):27-29
[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol... 相似文献
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Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed. 相似文献
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D. A. Bond G. J. Jellis G. G. Rowland J. Le Guen L. D. Robertson S. A. Khalil L. Li-Juan 《Euphytica》1993,73(1-2):151-166
Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses. 相似文献
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Chunli TANG Fengxian ZHAO Hongxia CHEN Jiabao MA Jiangcun WEI Meiyan QIU Zhen XIE 《Medicinal Plant》2019,(6):60-65
[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination... 相似文献
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E. Keep 《Euphytica》1986,35(3):843-855
Summary Cytoplasmic male sterility (cms) is described in the F1 hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf
1) and recessive for the other (Rf
2).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph
3(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf
1(one of two dominant additive genes controlling early season leafing out) indicated that Rf
1and Rf
2were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf
1, and 0.15 for Lf
1-Sph
3were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph
3.It is suggested that competitive disadvantage of lf
1-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph
3. Poor performance of lf
1- (and possibly lf
2-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off). 相似文献
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[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho... 相似文献
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Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed. 相似文献
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Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques. 相似文献