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A method was developed for high resolution electrophoresis of proteins in linear gradient (3 to 30%) polyacrylamide gel rods in a neutral phosphate buffer containing 0.1% sodium dodecyl sulfate. Well-defined protein zones were observed and improved resolution was attained especially for low molecular weight proteins in preparations containing a variety of polypeptides, e. g. viruses that are often separated by continuous gel methods. Electropherograms of continuous (8%)and gradient (3 to 30%) gels were made of purified vesicular stomatitis virus, variola virus, Rickettsia rickett-sii, and alpha and beta chains of hemoglobin in order to demonstrate the resolution of the gradient system. 相似文献
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Seiichi Nasuno 《Bioscience, biotechnology, and biochemistry》2013,77(7):1147-1150
Ciliatine (2-aminoethylphosphonic acid) (76 mg) was isolated from 72 g of lipids of the oyster with a combination of ion exchange chromatographic techniques and was identified from the results of elementary analysis, infrared spectrum, and chromatographic behaviors. The phosphonic acid was also detected in hydrolysates of a chloroform-methanol insoluble fraction of the oyster. It has been demonstrated that the oyster contains high concentration of ciliatine. 相似文献
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Plant Nucleases: III. Polyacrylamide Gel Electrophoresis of Corn Ribonuclease Isoenzymes 总被引:1,自引:1,他引:0 下载免费PDF全文
Wilson CM 《Plant physiology》1971,48(1):64-68
Isoenzymes of RNase were detected in plant extracts after polyacrylamide gel electrophoresis with a new buffer system. The gels were incubated in an RNA solution, then dipped for 30 seconds into 0.2% toluidine blue. The method is rapid and is sensitive to very small amounts of RNase. The effects of buffers and ethylenediaminetetraacetate on the different enzymes are illustrated by photographs and scans of the gels. 相似文献
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Urea PAGE or denaturing urea polyacrylamide gel electrophoresis employs 6-8 M urea, which denatures secondary DNA or RNA structures and is used for their separation in a polyacrylamide gel matrix based on the molecular weight. Fragments between 2 to 500 bases, with length differences as small as a single nucleotide, can be separated using this method1. The migration of the sample is dependent on the chosen acrylamide concentration. A higher percentage of polyacrylamide resolves lower molecular weight fragments. The combination of urea and temperatures of 45-55 °C during the gel run allows for the separation of unstructured DNA or RNA molecules.In general this method is required to analyze or purify single stranded DNA or RNA fragments, such as synthesized or labeled oligonucleotides or products from enzymatic cleavage reactions.In this video article we show how to prepare and run the denaturing urea polyacrylamide gels. Technical tips are included, in addition to the original protocol 1,2. 相似文献
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Shun-Jen Tsai Michiko Yamashita Soichi Arai Masao Fujimaki 《Bioscience, biotechnology, and biochemistry》2013,77(3):641-644
A peptic hydrolysate of soybean protein was filtered with Sephadex G–25 and was separated approximately into four fractions (I, II, II, and IV in the order of mol. wt.). Fraction II (av. mol. wt: 1043) and III (av. mol. Wt.: 685) were more plastein-productive than others. When plastein produced from Fraction II with Nagarse was investigated by plate electrophoresis using 7.5% polyacrylamide-gel, the upper limit of the molecular weight was found to be about 25,000. A similar result was obtained also with Fraction III. The increase of molecular weight in the course of the plastein formation with the mixture (substrate) of Fractions II and III was shown that the final product lay mainly in a position between cytochrome c (mol. wt.: 11,700) and Nagarse (mol wt.: 27,600). In addition, the gel-electrophoretic experiments revealed that the most favorable condition for the plastein synthesis were pH 6.5 and 35% in substrate concentration. 相似文献
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High-resolution polyacrylamide gradient gel electrophoresis (PGGE) was used to separate isoenzymes of 12 Naegleria strains: one N. australiensis, two N. lovaniensis, one N. jadini, two N. gruberi isolated from environmental samples, and six N. fowleri strains isolated from patients with primary amoebic meningoencephalitis. Of the eight enzymes studied, seven showed zymograms with interspecific variation that identified all the species tested. Although the six N. fowleri strains were biochemically the most homogeneous, they showed intraspecific isoenzyme variation that allowed them to be grouped into four zymodemes. The PGGE technique, which separates isoenzymes by their molecular shape, is both sensitive and economical. It offers an addition or an attractive alternative to isoelectric focusing which has commonly been used to aid species identification of Naegleria by separating isoenzymes by their isoelectric point. 相似文献
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High-Resolution Differentiation of Cyanobacteria by Using rRNA-Internal Transcribed Spacer Denaturing Gradient Gel Electrophoresis 总被引:1,自引:1,他引:0 下载免费PDF全文
Ingmar Janse Marion Meima W. Edwin A. Kardinaal Gabriel Zwart 《Applied microbiology》2003,69(11):6634-6643
For many ecological studies of cyanobacteria, it is essential that closely related species or strains can be discriminated. Since this is often not possible by using morphological features, cyanobacteria are frequently studied by using DNA-based methods. A powerful method for analysis of the diversity and dynamics of microbial populations and for checking the purity and affiliation of cultivated strains is denaturing gradient gel electrophoresis (DGGE). We realized high-resolution discrimination of a variety of cyanobacteria by means of DGGE analysis of sections of the internal transcribed spacer between the 16S and 23S rRNA genes (rRNA-ITS). A forward primer specific for cyanobacteria, targeted at the 3′ end of the 16S rRNA gene, was designed. The combination of this primer and three different reverse primers targeted to the rRNA-ITS or to the 23S rRNA gene yielded PCR products of different sizes from cultures of all 16 cyanobacterial genera that were tested but not from other bacteria. DGGE profiles produced from the shortest section of rRNA-ITS consisted of one band for all but one cyanobacterial genera, and those generated from longer stretches of rRNA-ITS yielded DGGE profiles containing one to four bands. The suitability of DGGE for detecting intrageneric and intraspecific variation was tested by using strains of the genus Microcystis. Many strains could be discriminated by means of rRNA-ITS DGGE, and the resolution of this method was strikingly higher than that obtained with previously described methods. The applicability of the developed DGGE assays for analysis of cyanobacteria in field samples was demonstrated by using samples from freshwater lakes. The advantages and disadvantages associated with the use of each developed primer set are discussed. 相似文献
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LDS-聚丙烯酰胺凝胶电泳分析植物过氧化物酶同工酶 总被引:4,自引:0,他引:4
以高粱和甘蓝型油菜叶片为材料,用十二烷基磺酸锂不连续聚丙烯酰胺凝胶电泳(LDS-PAGE)检测过氧化物酶同工酶的结果表明,LDS-PAGE的酶带条数比非变性不连续聚丙烯酰胺凝胶电泳(native PAGE)明显增加,分辨率大大提高,上样量少,胶片易保存;比复性电泳(G-PAGE)的步骤简单,电泳后无需除去LDS即可直接按常规活性染色方法染色。 相似文献
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DGGE/TGGE技术及其在微生物分子生态学中的应用 总被引:48,自引:1,他引:48
变性梯度凝胶电泳(DGGE)和温度梯度凝胶电泳(TGGE)是近些年微生物分子生态学研究中的热点技术之一。由于DGGE/TGGE技术具有可靠性强、重现性高、方便快捷等优点,被广泛地应用于微生物群落多样性和动态性分析。文章对DGGE/TGGE技术原理与关键环节、局限性和应用前景进行了综述。 相似文献
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S ummary : Colicins can be separated by polyacrylamide gel electrophoresis, so allowing multiple colicinogeny to be detected with a universal indicator strain. 相似文献
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The flat gel acrylamide electrophoresis technique was standardized and applied to the comparison of four species of porcine mycoplasmas. Clear differences were observed between these species, and differences were seen among the strains of Mycoplasma hyosynoviae. The clarity of the patterns and the number of bands developed was influenced by the amount of protein in the extract and the age of the culture. The technique allows the comparison of several protein extracts in parallel without the problems associated with the rearrangement of separate gel columns. 相似文献
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王瑞珍 《中国生物化学与分子生物学报》1991,7(1):79-82
用不连续SDS-聚丙烯酰胺凝胶电泳分析了人脑胶质细胞瘤与正常脑细胞核NHCP电泳图谱。从二者的NHCP电泳结果表明,脑胶质细胞瘤增加了一条表观分子量为3万的蛋白区带;表观分子量为1.75万~6.5万的蛋白区带染色明显加深。染色质紫外吸收光谱也有明显差异。总之,脑胶质细胞瘤核NHCP发生质与量的变化。 相似文献
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应激心肌细胞蛋白质组双向凝胶电泳分析 总被引:14,自引:0,他引:14
采用双向凝胶电泳技术和计算机辅助的图像分析方法 ,对去甲肾上腺素诱导的应激心肌细胞与正常心肌细胞蛋白质进行分离和比较分析 .正常心肌细胞可分离 12 32± 5 6个蛋白点 ,蛋白点匹配率为 83 3%± 1 0 %.有 11种蛋白质在NE应激后发生了明显和稳定的质和量的改变 (P <0 0 5 ) ,其中 6种 (Mr pI :4 9 7kD 7 8,38 3kD 5 9,37 1kD 6 6 ,2 9 3kD 7 4 ,18 7kD 6 1,18 5kD 7 7)在应激后表达降低 ,4种 (Mr pI:4 7 6kD 5 5 ,31 9kD 4 4 ,2 6 6kD 4 6 ,33 2kD 8 1)在应激后表达增高 ,1种 (Mr pI:19 4kD 6 9)只在应激后发生表达 .这些差异表达的蛋白质可能参与了心血管应激反应乃至应激损伤发生的过程 . 相似文献
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Identification of Frankia Strains by Two-Dimensional Polyacrylamide Gel Electrophoresis 总被引:4,自引:3,他引:1 下载免费PDF全文
Fifteen Frankia strains from five different plant species were analyzed by two-dimensional polyacryl-amide gel electrophoresis to determine their relatedness by comparing the polypeptide patterns obtained. Three major subgroups (A, C, and D) were found in the Alnus-Comptonia-Myrica cross-inoculation group. An isolate from Purshia tridentata had a unique protein pattern and represents a distinct group of frankiae. Members of group A were isolated from root nodules of Alnus incana subsp. rugosa and Alnus viridis subsp. crispa. Group C organisms were from A. incana subsp. rugosa and Comptonia peregrina nodules, and group D organisms were from A. incana subsp. rugosa, A. viridis subsp. cripsa, and Myrica pensylvanica root nodules. Isolates from each gel group were obtained at several widely separated geographical locations. The results indicate that two-dimensional polyacrylamide gel electrophoresis is useful for identifying Frankia isolates. 相似文献
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Combination of Multiplex PCR and PCR-Denaturing Gradient Gel Electrophoresis for Monitoring Common Sourdough-Associated Lactobacillus Species 下载免费PDF全文
Luca Settanni Sara Valmorri Douwe van Sinderen Giovanna Suzzi Antonello Paparella Aldo Corsetti 《Applied microbiology》2006,72(5):3793-3796
A combination of denaturing gradient gel electrophoresis (DGGE) and a previously described multiplex PCR approach was employed to detect sourdough lactobacilli. Primers specific for certain groups of Lactobacillus spp. were used to amplify fragments, which were analyzed by DGGE. DGGE profiles obtained from Lactobacillus type strains acted as standards to analyze lactobacilli from four regional Abruzzo (central Italy) sourdoughs. 相似文献