共查询到20条相似文献，搜索用时 125 毫秒
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本文就植物基因组原位杂交中出现的无杂交信号，杂交信号过多（杂交背景重）或过少，染色体丢失及杂交污点产生的原因进行了初步分析，并提出了一些解决的方法。 相似文献
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利用放射免疫技术快速检测转基因植物 总被引：2，自引：0，他引：2
利用PCR分别合成掺入地高辛标记的35S启动子和NOS终止子核酸探针，再利用放射免疫技术将I^125标记的地高辛抗体与35S启动子和NOS终止子探针分别进行杂交检测。结果显示转基因样本的杂交信号均为阳性，且灵敏度较高。特异性强，为转基因植物的分析检测又提供了一种较实用的方法。 相似文献
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朱来华;梁成珠;陆承平;吴华;杨元杰;马丰忠;王树峰;于红光 《农业生物技术学报》2006,14(2):203-207
通过分子克隆技术获得马疱疹病毒1型(Equine herpesvirus 1, EHV1)、马动脉炎病毒(Equine arteritis virus , EAV)、马流感病毒(Equine influenza virus, EIV)、马传染性贫血病毒(Equine infectious anaemia virus , EIAV)和东部马脑脊髓炎病毒(Eastern equine encephalomyelitis virus, EEEV)等5种病毒各一段高度保守的特异性基因片段，用芯片点样仪逐点分配到处理过的玻片上，制备成检测芯片。提取样品中的RNA，进行反转录和荧光标记后滴加到芯片上进行特异性杂交，对杂交结果进行扫描检测和计算机软件分析。结果显示，制备的基因芯片可同时检测和鉴别上述5种病毒，可检测到阳性杂交信号的最高稀释度为10-6的病毒液，约25个病毒DNA拷贝，但其它病毒材料未见红色荧光信号，证明了本方法的特异性。在进口马的隔离检疫期间，采集马鼻肺炎、马动脉炎中和抗体阳性但病毒分离阴性马匹的白细胞悬液，分别在EHV1和EAV位点处可检测到阳性杂交信号。证明基因芯片技术不但快速、准确和敏感，而且可同时进行多种病毒的检测。 相似文献
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酵母双杂交系统的研究进展 总被引：7，自引：0，他引：7
酵母双杂交系统是研究蛋白质间相互作用的一种分子生物学方法，随着该系统的广泛应用，发展了三杂交系统、单杂交系统，及其它研究蛋白质与DNA、RNA相互作用的方法。该方法在研究蛋白质组学领域有重要地位和作用。 相似文献
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基因芯片技术检测5种马病毒 总被引：8，自引：0，他引：8
通过分子克隆技术获得马疱疹病毒1型(Equineherpesvirus1,EHV1)、马动脉炎病毒(Equinearteritisvirus,EAV)、马流感病毒(Equineinfluenzavirus,EIV)、马传染性贫血病毒(Equineinfectiousanaemiavirus,EIAV)和东部马脑脊髓炎病毒(East-ernequineencephalomyelitisvirus,EEEV)等5种病毒各一段高度保守的特异性基因片段,用芯片点样仪逐点分配到处理过的玻片上,制备成检测芯片。提取样品中的RNA,进行反转录和荧光标记后滴加到芯片上进行特异性杂交,对杂交结果进行扫描检测和计算机软件分析。结果显示,制备的基因芯片可同时检测和鉴别上述5种病毒,可检测到阳性杂交信号的最高稀释度为10-6的病毒液,约25个病毒DNA拷贝,但其它病毒材料未见红色荧光信号,证明了本方法的特异性。在进口马的隔离检疫期间,采集马鼻肺炎、马动脉炎中和抗体阳性但病毒分离阴性马匹的白细胞悬液,分别在EHV1和EAV位点处可检测到阳性杂交信号。证明基因芯片技术不但快速、准确和敏感,而且可同时进行多种病毒的检测。 相似文献
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杂交水稻机械化种子生产技术的研究进展 总被引：5，自引：4，他引：1
该文回顾分析了国内外杂交水稻制种技术的发展概况,提出中国杂交水稻种子生产技术经历了杂交水稻制种繁种的经验摸索和积累阶段（1973～1980年）、杂交水稻制（繁）种的技术提高和完善阶段（1981～1990年）和杂交水稻制（繁）种的超高产技术研究与推广阶段（1991～至今）等3个发展阶段;国外杂交水稻大面积推广应用与与制种技术研究还在进行之中。认为要提高中国杂交水稻种子质量和种子生产效益及种子生产技术竞争力,今后必须应用推广杂交水稻机械化种子生产技术。至今已进行了除草剂敏感基因导入父本法、抗除草剂基因导入母本法和稻壳颜色标记法的杂交水稻混播机械化制种技术;杂交水稻机械采粉贮粉与授粉制种技术;利用雌性不育系的机械制种技术等。其中杂交水稻混播机械化制种技术已获得小面积成功应用,其余方法还在试验与研究之中。因此,今后需要水稻育种家与农机专家密切合作,进一步深入开展杂交水稻机械化制种技术的研究及推广应用工作。 相似文献
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基因芯片是以预先设计的方式将大量的生物讯息密码（寡核苷酸、cDNA、基因组DNA等）固定在玻片、硅片等固相载体上组成的密集分子阵列。基因芯片技术本质是生物信号的平行分析,它利用核酸分子杂交原理,通过荧光标记技术检测杂交亲和与否,再经过计算机分析处理可迅速获得所需信息。由于其具有高通量、微型化、连续化、自动化、快速和准确等特点,已引起国际国内广泛的关注和重视,在许多领域得到了广泛的应用。本文简述了基因芯片的概念,技术特点及主要分类,着重对其在基因表达水平检测,基因突变和多态性的分析,基因组DNA分析,后基因组学研究以及转基因农作物检测等方面进行阐述,并说明其存在的问题及展望。 相似文献
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从大田稻株上采取杂交适期穗，室内进行去雄，授粉等人工杂交后，将杂交插在水中经４０天管理即可获得杂交种子，应用本杂交法，杂交适期穗可根据所需数量任意采取，省去了母本移栽手续，有操作方便，管理简单等优点。 相似文献
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为发掘陆地棉纤维品质性状基因/数量性状基因座(quantitative trait loci,QTL),本研究以陆地棉(Gossypium hirsutum L.)优质品种渝棉1号分别与贝尔斯诺和中棉所35杂交获得F1代,随后利用两个F1再次杂交得到复合杂交群体.利用6565对SSR引物筛选渝棉1号、中棉所35和贝尔斯诺,获得155对多态性引物.对检测[(渝棉1号×中棉所35)x(渝棉1号×贝尔斯诺)]F1复合杂交群体标记基因型获得的158个位点进行遗传连锁分析,构建的遗传图谱包含102个位点,覆盖1199.0 cM.利用多QTL(multiple-QTL model,MQM)作图法,检测到11个纤维品质性状QTL,包括4个纤维长度、2个长度整齐度和5个纤维比强度QTL.11个QTL分布于4条D基因组染色体.本研究表明,陆地棉不同品种同一纤维品质性状QTL等位基因存在差异;三亲本杂交群体不仅可以提高群体多态性分子标记比例,而且可以显示同一QTL在不同遗传背景的等位基因表现差异. 相似文献
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S. Feng R. D. Wolfinger T. M. Chu G. C. Gibson L. A. McGraw 《Journal of Agricultural, Biological & Environmental Statistics》2006,11(2):197-209
A gene-by-gene mixed model analysis is a useful statistical method for assessing significance for microarray gene differential
expression. While a large amount of data on thousands of genes are collected in a microarray experiment, the sample size for
each gene is usually small, which could limit the statistical power of this analysis. In this report, we introduce an empirical
Bayes (EB) approach for general variance component models applied to microarray data. Within a linear mixed model framework,
the restricted maximum likelihood (REML) estimates of variance components of each gene are adjusted by integrating information
on variance components estimated from all genes. The approach starts with a series of single-gene analyses. The estimated
variance components from each gene are transformed to the “ANOVA components”. This transformation makes it possible to independently
estimate the marginal distribution of each “ANOVA component.” The modes of the posterior distributions are estimated and inversely
transformed to compute the posterior estimates of the variance components. The EB statistic is constructed by replacing the
REML variance estimates with the EB variance estimates in the usual t statistic. The EB approach is illustrated with a real data example which compares the effects of five different genotypes
of male flies on post-mating gene expression in female flies. In a simulation study, the ROC curves are applied to compare
the EB statistic and two other statistics. The EB statistic was found to be the most powerful of the three. Though the null
distribution of the EB statistic is unknown, a t distribution may be used to provide conservative control of the false positive rate. 相似文献
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Rapid and reliable detection and identification of GM events using multiplex PCR coupled with oligonucleotide microarray 总被引：7，自引：0，他引：7
Xu X Li Y Zhao H Wen SY Wang SQ Huang J Huang KL Luo YB 《Journal of agricultural and food chemistry》2005,53(10):3789-3794
To devise a rapid and reliable method for the detection and identification of genetically modified (GM) events, we developed a multiplex polymerase chain reaction (PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a single reaction. The system included probes for screening gene, species reference gene, specific gene, construct-specific gene, event-specific gene, and internal and negative control genes. 18S rRNA was combined with species reference genes as internal controls to assess the efficiency of all reactions and to eliminate false negatives. Two sets of the multiplex PCR system were used to amplify four and five targets, respectively. Eight different structure genes could be detected and identified simultaneously for Roundup Ready soybean in a single microarray. The microarray specificity was validated by its ability to discriminate two GM maizes Bt176 and Bt11. The advantages of this method are its high specificity and greatly reduced false-positives and -negatives. The multiplex PCR coupled with microarray technology presented here is a rapid and reliable tool for the simultaneous detection of GM organism ingredients. 相似文献
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十字花科黑腐病菌RNA提取与质量鉴定 总被引：1，自引：0，他引：1
十字花科黑腐病菌,学名野油菜黄单胞菌野油菜致病变种（Xanthomonas campestris pv.campestris,简称Xcc）,是一类γ-变形菌纲的革兰氏阴性细菌,能在世界范围内侵染十字花科植物,给农业生产造成重大损失。RNA是分子生物学的主要研究对象之一,提取高质量的RNA是研究十字花科黑腐病菌基因表达调控机理的基础。由于Xcc能产生大量胞外多糖及黄单胞色素,且细菌RNA半衰期较短,目前尚缺少一种大量提取Xcc高质量RNA的有效方法。该研究在参考多种RNA提取方法的基础上,建立了一种简单、有效的适合十字花科黑腐病菌RNA的提取方法。得到的RNA样品经过紫外分光光度法和甲醛变性凝胶电泳检测,证实为完整、均一的总RNA,达到了表达谱分析及cDNA文库构建的要求。 相似文献
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Herv Sanguin Lionel Kroneisen Kevin Gazengel Martina Kyselkov Benoît Remenant Claire Prigent-Combaret Genevive L. Grundmann Alain Sarniguet Yvan Moënne-Loccoz 《Soil biology & biochemistry》2008,40(5):1028-1039
So far, the analysis of microbial populations associated with wheat monocropping-induced decline of take-all disease (Gaeumannomyces graminis var. tritici) has focused mainly on culturable biocontrol pseudomonads. The objective of this study was to develop a taxonomic rrs (16S rRNA gene) microarray to assess the changes in Pseudomonas populations taking place during take-all decline. The microarray contains 12 probes for five Pseudomonas phylogenetic clusters chosen because they include well-known plant-beneficial pseudomonads. Four of the clusters are within the ‘Pseudomonas fluorescens’ species complex. PCR primers were selected to target these five clusters, and they were validated using 53 pseudomonads belonging or not to these clusters. Microarray analysis of the pseudomonads enabled discrimination between strains from several Pseudomonas clusters. Rhizosphere samples were collected from field plots grown with wheat for 1 (low level of take-all disease), 5 (high level of disease) or 10 years (low level of disease, suppressiveness reached). Microarray data could distinguish Pseudomonas populations from some of the wheat plants grown in the same plot. When comparing treatments, there was a difference between years 1 and 10. Cloning–sequencing of rrs enabled to define more precisely this difference by identifying two major Pseudomonas populations, one associated with year 1 and the other with year 10 (disease suppressiveness), which represent new clades within the ‘P. fluorescens’ complex. These populations may be useful as soil quality indicators. In conclusion, the combination of microarray and cloning–sequencing approaches highlighted changes in the prevalence of two major Pseudomonas populations, giving new insights on the dynamics of root-associated pseudomonads during take-all decline. 相似文献
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Honda S Aoki F Tanaka H Kishida H Nishiyama T Okada S Matsumoto I Abe K Mae T 《Journal of agricultural and food chemistry》2006,54(24):9055-9062
Turmeric, the rhizome of Curcuma longa L., has a wide range of effects on human health. Turmeric oleoresin, an extract of turmeric, is often used for flavoring and coloring. Curcuminoids and turmeric essential oil are both contained in turmeric oleoresin, and both of these fractions have hypoglycemic effects. In the present study, we comprehensively assessed the effect of turmeric oleoresin on hepatic gene expression in obese diabetic KK-Ay mice using DNA microarray analysis and quantitative real-time polymerase chain reaction (PCR). Female KK-Ay mice aged 6 weeks (n = 6/group) were fed a high-fat diet containing turmeric oleoresin, curcuminoids, and essential oil for 5 weeks. The same diet without any of these fractions was used as a control diet. Ingestion of turmeric oleoresin and essential oil inhibited the development of increased blood glucose and abdominal fat mass, while curcuminoids only inhibited the increase in blood glucose. DNA microarray analysis indicated that turmeric oleoresin ingestion up-regulated the expression of genes related to glycolysis, beta-oxidation, and cholesterol metabolism in the liver of KK-Ay mice, while expression of gluconeogenesis-related genes was down-regulated. Real-time PCR analysis was conducted to assess the contribution of the curcuminoids and essential oil in turmeric oleoresin to the changes in expression of representative genes selected by DNA microarray analysis. This analysis suggested that curcuminoids regulated turmeric oleoresin ingestion-induced expression of glycolysis-related genes and also that curcuminoids and turmeric essential oil acted synergistically to regulate the peroxisomal beta-oxidation-related gene expression induced by turmeric oleoresin ingestion. These changes in gene expression were considered to be the mechanism by which the turmeric oleoresin affected the control of both blood glucose levels and abdominal adipose tissue masses. All of these results suggest that the use of whole turmeric oleoresin is more effective than the use of either curcuminoids or the essential oil alone. 相似文献
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Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray 总被引：5，自引：0，他引：5
Xu J Zhu S Miao H Huang W Qiu M Huang Y Fu X Li Y 《Journal of agricultural and food chemistry》2007,55(14):5575-5579
With the increasing development of genetically modified organism (GMO) detection techniques, the polymerase chain reaction (PCR) technique has been the mainstay for GMO detection. An oligonucleotide microarray is a glass chip to the surface of which an array of oligonucleotides was fixed as spots, each containing numerous copies of a sequence-specific probe that is complementary to a gene of interest. So it is used to detect ten or more targets synchronously. In this research, an event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity using multiplex-PCR together with oligonucleotide microarray. A commercial GM soybean (GTS 40-3-2) and six GM maize events (MON810, MON863, Bt176, Bt11, GA21, and T25) were detected by this method. The results indicate that it is a suitable method for the identification of these GM soybean and maizes. 相似文献
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Developments in soil microbiology since the mid 1960s 总被引：11，自引：0，他引：11
Heribert Insam 《Geoderma》2001,100(3-4):389-402
Since the 1960s, soil microbiology underwent major changes in methods and approaches and this review focuses on the developments in some selected aspects of soil microbiology. Research in cell numbers of specific bacterial and fungal groups was replaced by a focus on biochemical processes including soil enzyme activities, and flux measurements of carbon and nutrients. Ecologists focused on soil microbial pools whereas soil microbial biomass as an important source and sink of nutrients were recognized in agriculture. Soil microbiologists started to use structural components like phospholipid fatty acids for quantification of specific microbial groups without the need to cultivate them. In the last decade, molecular approaches allowed new insights through the analysis of soil extract DNA showing an unexpected diversity of genomes in soil. At the end of the review a brief outlook is given on the future of soil microbiology which ranges from in situ identification of bacteria, to routine assays of microbial communities by microarray technology. 相似文献
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针对目前通用的cDNA微阵列数据的标准化处理方法(常称为对数比转换法)中存在的缺陷，提出了一种新的cDNA微阵列数据标准化方法－非转换法。该方法利用Huber等(2003)提出的芯片数据分析模型，在对芯片背景进行校正后，根据最小二乘估计原理，通过迭代的方式直接对样品中的基因转录水平进行估计，从而达到数据标准化的目的。利用计算机模拟，比较了在各种条件下(不同的芯片内探针重复数、不同的背景变异程度、不同的差异表达基因上下调比例等)该方法与对数比转换法的优劣。结果表明，在所有条件下，非转换法都优于对数比转化法，由非转换法得到的估计值的相对偏差都在5％以下，而由对数比转化法得到的估计值的相对偏差都在20％以上，而且对数比转化法对背景变异和差异表达基因上下调比例更敏感，随着背景变异程度的增大和差异表达基因上下调比例不对称性的提高，其估计值的偏差急剧上升，而非转换法则基本不受影响。因而非转换法可以作为cDNA微阵列数据标准化的一种替代方法。 相似文献