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High-performance electrophoresis chromatography (HPEC) is a recent development that features continuous-elution gel electrophoresis for isolating proteins or peptides in range of 1 to 300 microgram quantities. Column gel electrophoresis is conducted under thermostated conditions, and the field voltage can be varied within a run with a programmable power supply. Applications of this apparatus in protein purification are presented to demonstrate the utility of the (Model 230A) HPEC. These examples include on-line detection with direct analyte recovery of highly purified sample, which mimics high-performance liquid chromatography, for subsequent structure-function characterization. A method to remove salts from sodium dodecyl sulfate electrophoresed samples for subsequent sequencing or amino acid analysis is described. This desalting procedure recovers from 90%-95% of the sample and employs a low molecular weight cut-off membrane during sample centrifugation onto a polyvinylidene difluoride membrane. Subsequent washings are performed to efficiently remove salts, free amino acids and detergents that are known to interfere with sequence analysis. Sequence information such as initial recovery, repetitive yields and chromatogram comparisons are presented to demonstrate the utility of this procedure when used following isolation of sample with HPEC. 相似文献
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Effect of Inorganic Phosphate on the Biosynthesis of Purine and Pyrimidine Nucleotides in Suspension-Cultured Cells of Catharanthus roseus 总被引:1,自引:0,他引:1
The levels of purine and pyrimidine nucleotides in suspensioncultures of Catharanthus roseus were determined 24 h after stationary-phasecells were transferred to fresh complete (+Pi)or phosphate-deficient (Pi) Murashige-Skoogmedium. The levels of ATP, GTP, UTP and CTP were from approx.3 to 5-fold greater in the cells grown in +Pimedium than in the cells grown in Pi medium.The levels of almost all other nucleotides were slightly higherin the cells in +Pi medium. The rates of de novoand salvage biosynthesis of purine and pyrimidine nucleotideswere estimated from the rates of incorporation of radioactivityfrom [14C]formate, [214C]glycine, NaH14CO3, [614C]orotate,[814C]adenine, [814C]adenosine, [214C]uraciland [214C]uridine. The results indicated that the activityof both the de novo and the salvage pathway was higher in thecells in +Pi medium than in the cells in Pimedium. The rate of degradation estimated from the rate of releaseof 14CO2 from labelled purines and pyrimidines indicated thatdegradation of uridine was significantly reduced in the cellsin +Pi medium, but no significant difference wasfound in the degradation of adenine, adenosine and uracil. Thepossible role of Pi in the control of the biosynthesis of nucleotidesand in the degradation of uridine is discussed. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, nucleotides, purines, pyrimidines, biosynthesis, degradation 相似文献
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电针、吗啡镇痛和耐受时某些脑区线粒体结合钙的变化 总被引:1,自引:0,他引:1
采用 Tb~(3+)荧光探针和离子选择电极研究了电针和吗啡镇痛及镇痛耐受时,动物不同脑区游离 Ca~(2+)和线粒体膜结合 Ca~(2+)的变化。实验结果表明,电针和吗啡都有较强的镇痛作用,与此同时,导水管周围灰质和下丘脑的线粒体膜结合 Ca~(2+)升高。脑室内预注钌红,则能降低这两个脑区的线粒体膜结合 Ca~(2+)和痛阈。另一方面,在电针或吗啡耐受时,两脑区的游离 Ca~(2+)浓度增加,线粒体膜结合 Ca~(2+)降低。结果提示,神经细胞质膜内外 Ca~(2+)的移动可能在电针和吗啡镇痛中起某种调节作用。 相似文献
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柱霉属一新种——蝙蝠蛾柱霉 总被引:5,自引:0,他引:5
从青海省化隆地区采集冬虫夏草新鲜标本,经分离培养获一纯种,并经鉴定是隶属暗色孢科Dematiaceae的柱霉属Scytalidium Pesante,其形态特征与近似种——黄棕柱霉Scytalidium fulvum进行了比较,由于分生孢子量度及生活习性与黄棕柱霉有着明显差别,故定为一新种,命名为蝙蝠蛾柱霉Scytalidium hepiali C.L.Li sp.nov.该新种在PDA培养基上生长迅速,25℃,培养10天菌落直径2.5cm。菌丝相互平排列成菌丝束。产孢细胞裂殖产孢,节孢子2型:(1)无色,透明,薄壁,圆柱形,桶形。(2)淡褐色,厚壁,椭圆形,桶形,亚球形。蝙蝠蛾柱霉在蛋白胨、葡萄糖液体培养基中进行深层培养,26℃,20天。发酵液用乙醇抽提,减压蒸馏,甲醇溶解,过滤,减压浓缩,蒸馏水溶解,冷冻干燥。从1升发酵液中获虫草菌素制品50~70mg。从拮抗试验表明虫草菌素能抑制枯草芽孢杆菌的生长。 相似文献
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本文利用含有抗核基质自发抗体的硬皮人血清,以小鼠艾氏腹水癌细胞为材料,用间接免疫荧光染色的方法,追踪了对应核基质抗原在细胞周期中分布的变化。结果显示,在末期和间期之间存在一个核基质抗原从细胞质向细胞核内转移的过程。由于这一过程是通过核膜进行的,从而提示核基质结构可能有解聚和再聚合的行为。用酶化学结合间接免疫荧光染色的方法,初步研究了抗原的化学性质。染色形态的比较研究显示所用血清中可能含有不同于以前发现的、抗新的核基质抗原的自发抗体。 相似文献