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从固体平板挑取转化带有目的基因的单菌落 ,用特异引物通过聚合酶链式反应可直接扩增和标记目的基因 ,不需经过菌的液体培养、质粒提取和酶解反应等复杂过程 ,能快速获得目的基因扩增产物和进行目的基因探针的标记。 相似文献
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The phytohormone salicylic acid(SA) regulates biotic and abiotic stress responses in plants. Two distinct biosynthetic pathways for SA have been well documented in plants: the isochorismate(IC)pathway in the chloroplast and the phenylalanine ammonia-lyase(PAL) pathway in the cytosol.However, there has been no solid evidence that the PAL pathway contributes to SA biosynthesis. Here,we report that feeding Arabidopsis thaliana with Ring-13C-labeled phenylalanine(13C6 相似文献
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本文报道了以人绒毛细胞为材料, 利用人类Y染色体特异片段的一对扩增引物Y~3|和Y~4|以地高辛(Digoxigenin)为标记物, 在玻片上进行细胞内引物原位延伸标记,进而快速检测Y染色体专性序列的实验结果。Abstract:By using primers that generate DNA segments specific for the human Y chromosome,we have successfully carried out primed in situ Digoxigenin labeling at single-cell level on the glass slides.The extreme speed,simplicity and safey make it possible to be used to detect sex and diagnosis chromosome disorder. 相似文献
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Huang WE Huang L Preston GM Naylor M Carr JP Li Y Singer AC Whiteley AS Wang H 《The Plant journal : for cell and molecular biology》2006,46(6):1073-1083
Salicylic acid (SA) plays important roles in plants, most notably in the induction of systemic acquired resistance (SAR) against pathogens. A non-destructive in situ assay for SA would provide new insights into the functions of SA in SAR and other SA-regulated phenomena. We assessed a genetically engineered strain of Acinetobacter sp. ADP1, which proportionally produces bioluminescence in response to salicylates including SA and methylsalicylate, as a reporter for salicylate accumulation in the apoplast of plant leaves. SA was measured quantitatively in situ in NN genotype tobacco (Nicotiana tabacum L. cv Xanthi-nc) leaves inoculated with tobacco mosaic virus (TMV). The biosensor revealed accumulation of apoplastic SA before the visible appearance of hypersensitive response (HR) lesions. When the biosensor was infiltrated into TMV-inoculated leaves displaying HR lesions at 90 and 168 h post-inoculation, salicylate accumulation was detected predominantly in tissues surrounding the lesions and in veins adjacent to HR lesions. These images are consistent with previous data demonstrating that SA accumulation occurs prior to and following the onset of visible HR lesions. We also used the biosensor to observe apoplastic SA accumulation in tobacco leaves inoculated with virulent and HR-eliciting strains of the bacterial plant pathogen Pseudomonas syringae. The work demonstrates that the Acinetobacter sp. ADP1 biosensor is a useful new tool to non-destructively assay salicylates in situ and to map their spatial distribution in plant tissues. 相似文献
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洞门遗址位于山西省襄汾县新城镇沙女沟村东1 km处,西距丁村遗址5.1 km。2015年10月~2016年5月,山西省考古研究所进行了为期4个月的发掘,发掘面积27 m2。遗址地层自上而下依次为表土层、马兰黄土层(L1)、棕红色古土壤层(S1)。包括石制品、动物化石碎片、炭屑等在内的文化遗物均出土于S1中。另外在遗址西侧100 m左右的L1内采集石制品2件。石制品原料以角页岩为主,类型包括石核、石片、石器、断块等。与此同时,在沙女沟村东黄土台塬的S1地层中发现多个类似的原地埋藏的石器地点,说明在末次间冰期丁村遗址群附近汾河东岸至大崮堆山之间的黄土塬区人类活动十分频繁,且持续较长时间。研究表明,洞门遗址是一个别于丁村遗址的河流相埋藏类型的临时营地,它的发现为进一步认识丁村人的活动范围、行为模式以及末次间冰期及末次冰期古人类活动提供了重要资料。 相似文献
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[目的]分离鉴定植物乳杆菌PC518的质粒并分析滚环复制p C194家族复制起点特征。[方法]从植物乳杆菌PC518中提取质粒,HindⅢ单酶切后克隆测序,然后用反向PCR方法验证质粒序列的完整性。使用DNAMAN V6. 0软件和MEGA X软件对43个p C194家族质粒的复制起点序列和复制蛋白进行比对分析。[结果]分离得到一个3 325 bp的新质粒p LP325。43个p C194家族质粒复制起点中:24个在nick上、下游均有反向重复序列,12个只在nick上游有反向重复序列,4个只在下游有反向重复序列。复制蛋白的聚类与复制起点中反向重复序列的位置是对应的。[结论]p LP325的复制方式推定为滚环复制,属于p C194家族。p C194家族复制起点的bind以反向重复序列为特征,位于nick上游或下游。 相似文献
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Although the number of phylotypes present in a microbial community may number in the hundreds or more, until recently, fluorescence in situ hybridization has been used to label, at most, only a handful of different phylotypes in a single sample. We recently developed a technique, CLASI-FISH for combinatorial labeling and spectral imaging – fluorescence in situ hybridization, to greatly expand the number of distinguishable taxa in a single FISH experiment. The CLASI technique involves labeling microbes of interest with combinations of probes coupled with spectral imaging to allow the use of fluorophores with highly overlapping excitation and emission spectra. Here, we present the basic principles and theory of CLASI-FISH along with some guidelines for performing CLASI-FISH experiments. We further include a protocol for creating fluorescence spectral reference standards, a vital component of successful CLASI-FISH. 相似文献
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Shi P Li D Chen H Xiong Y Wang Y Tian C 《Protein science : a publication of the Protein Society》2012,21(4):596-600
In this report, (19)F spin incorporation in a specific site of a specific membrane protein in E. coli was accomplished via trifluoromethyl-phenylalanine ((19) F-tfmF). Site-specific (19)F chemical shifts and longitudinal relaxation times of diacylglycerol kinase (DAGK), an E. coli membrane protein, were measured in its native membrane using in situ magic angle spinning (MAS) solid state nuclear magnetic resonance (NMR). Comparing with solution NMR data of the purified DAGK in detergent micelles, the in situ MAS-NMR data illustrated that (19)F chemical shift values of residues at different membrane protein locations were influenced by interactions between membrane proteins and their surrounding lipid or lipid mimic environments, while (19)F side chain longitudinal relaxation values were probably affected by different interactions of DAGK with planar lipid bilayer versus globular detergent micelles. 相似文献
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Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants 总被引:52,自引:0,他引:52
Tissa Senaratna Darren Touchell Eric Bunn Kingsley Dixon 《Plant Growth Regulation》2000,30(2):157-161
The hypothesis that physiologically activeconcentrations of salicylic acid (SA) and itsderivatives can confer stress tolerance in plants wasevaluated using bean (Phaseolus vulgaris L.) andtomato (Lycopersicon esculentum L.). Plantsgrown from seeds imbibed in aqueous solutions (0.1--0.5 mM) of salicylic acid or acetyl salicylic acid(ASA) displayed enhanced tolerance to heat, chillingand drought stresses. Seedlings acquired similarstress tolerance when SA or ASA treatments wereapplied as soil drenches. The fact that seedimbibition with SA or ASA confers stress tolerance inplants is more consistent with a signaling role ofthese molecules, leading to the expression oftolerance rather than a direct effect. Induction ofmultiple stress tolerance in plants by exogenousapplication of SA and its derivatives may have asignificant practical application in agriculture,horticulture and forestry. 相似文献
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黄鳝二价体上Px基因E3外显子特异引物原位DNA合成 总被引:1,自引:1,他引:1
在鱼类中首次应用地高辛标记的特异引物原位DNA合成技术,将Px基因E3外显子定位于黄鳝粗线期8号二价体的8q15~q26区间和l号二价体的lq32~q37区间,证实了异种生物基因探针和引物原位DNA合成技术用于基因定位研究的可行性。 相似文献
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Abstract a Micrococcus sp. isolated by isophthalate enrichment, utilized 8 of the 13 substituted benzoic acids tested as the sole source of carbon and energy. The organism degraded benzoic acid and anthranilic acid through the intermediate formation of catechol. While salicylate is metabolized through genetisic acid, p -hydroxybenzoic acid is degraded through protocatechuic acid. The organism grew well on isophthalate but failed to utilize phthalate and terphthalate. Catechol disoxygenase, gentisate dioxygenase and protocatechuate dioxygenase activities were shown in the cell-free extracts. Catechol and protocatechuate are further metabolized through an ortho -cleavage pathway. 相似文献
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The chelating agents, EDDHA, its iron salt, EDTA, and salicylic acid enhance bud formation in Bartramidula bartramioides (Griff.) Wijk & Marg. Salicylic acid elicits optimal response at 10–4 M , whereas the other substances do so at 10–7 M . Increased concentration of ferric citrate and cupric sulphate also stimulate bud induction. The accumulation of Fe3+ and Cu2+ is facilitated by chelators. The endogenous iron content is maximum at 10–7 M EDDHA or EDTA, which is also the concentration optimal for bud induction. 相似文献
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JAVIER SÁNCHEZ‐MARTÍN JIM HEALD ALISON KINGSTON‐SMITH ANA WINTERS DIEGO RUBIALES MARILUZ SANZ LUIS A. J. MUR ELENA PRATS 《Plant, cell & environment》2015,38(7):1434-1452
Although a wealth of information is available on the induction of one or several drought‐related responses in different species, little is known of how their timing, modulation and crucially integration influence drought tolerance. Based upon metabolomic changes in oat (Avena sativa L.), we have defined key processes involved in drought tolerance. During a time course of increasing water deficit, metabolites from leaf samples were profiled using direct infusion–electrospray mass spectroscopy (DI‐ESI‐MS) and high‐performance liquid chromatography (HPLC) ESI‐MS/MS and analysed using principal component analysis (PCA) and discriminant function analysis (DFA). The involvement of metabolite pathways was confirmed through targeted assays of key metabolites and physiological experiments. We demonstrate an early accumulation of salicylic acid (SA) influencing stomatal opening, photorespiration and antioxidant defences before any change in the relative water content. These changes are likely to maintain plant water status, with any photoinhibitory effect being counteracted by an efficient antioxidant capacity, thereby representing an integrated mechanism of drought tolerance in oats. We also discuss these changes in relation to those engaged at later points, consequence of the different water status in susceptible and resistant genotypes. 相似文献
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本文以大肠癌裸鼠移植瘤为动物模型,将实验分为双歧杆菌注射组以及肿瘤对照组,用原位末端标记法观察了移植瘤组织的凋亡细胞百分率。结果显示:双歧杆菌注射组大肠癌移植瘤的凋亡细胞百分率为26428±1862,而对照组则为2062±880,两者比较具有显著的统计学意义(P<0001)。提示青春型双歧杆菌可诱导大肠癌组织的凋亡,这可能是其抑瘤机理的一个方面 相似文献
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采用原位熔融共聚法,以淀粉和乳酸为原料,制备改性淀粉聚乳酸接枝共聚物,对共聚反应机理进行初步探讨.结果表明,淀粉经PEG-400和马来酸酐增塑改性处理反应活性增强,与乳酸反应生成的淀粉聚乳酸接枝共聚物分子量Mw为6.921×10~4,Mn为4.789×10~4,分子量分布Mw/Mn为1.445.共聚物在PBS缓冲溶液中进行降解测试,结果为6天后质量损失一半. 相似文献
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The effects of the treatment of different tissues of three bean cultivars (Pinto, Vilmorin and Arroz) with ultra‐violet (UV) UV‐B and UV‐C radiation and red light on the activity, quantity and RNA levels of NADP‐malic enzyme (NADP‐ME) were determined. Exposure to UV‐B radiation for 8 h caused a marked increase of NADP‐ME from leaves, stems and roots in the three cultivars studied. A similar induction was observed in the leaves and stems after 8 h of exposure under UV‐C, but not in the roots, suggesting that a different signal might be acting to induce the expression of NADP‐ME after UV‐B and UV‐C exposure. In contrast, red light was ineffective in inducing NADP‐ME in either tissue, so the regulation of the expression of this enzyme is phytocrome‐independent. The activity of superoxide dismutase, ascorbate peroxidase, catalase and peroxidase was also different in plants treated with UV‐B, UV‐C and photosynthetically active radiation, suggesting that various pathways may be acting in the regulation of these enzymes by UV‐B and UV‐C. Reactive oxygen species (ROS) were also required for UV‐B induction of NADP‐ME, as the addition of ascorbic acid before UV‐B treatment prevented NADP‐ME induction, whereas salicylic acid was not effective in inducing the enzyme, showing that NADP‐ME induction by UV‐B is ROS dependent but salicylic acid independent. 相似文献
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Altering the cell permeability by treating Cryptococcus neoformans with 1% (v/v) hexane stimulated the yield of transformation of n-pentadecane to the corresponding dioic acid, tridecane 1,13-dicarboxylic acid (DC-15); however, the biotransformation process was inhibited by the elevated levels of DC-15. To avoid product inhibition, a continuous process with immobilized cells was performed, and the result showed that the yield of DC-15 production was increased up to fivefold as compared with the batch type of DC-15 production. To integrate the product recovery process with the biotransformation, Amberlite XAD-2 resin was used for adsorbing DC-15 and configured as an external in situ product recovery system. The continuous process described in this study is adaptable for large-scale production of DC-15. 相似文献