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191.
192.
抗白粉病小麦染色体组型的分子标记与生化标记分析 总被引:1,自引:0,他引:1
应用与小麦第六同源群有关的分子和生化标记,包括DNA探针pSc5·3H3和pSR167以及同工酶Est-5和a-Amy-1,对来自六倍体小黑麦Beagle与普通小麦科冬58杂交后代F1花粉植株的抗白粉病株系M24.M09及M17进行了分析。结果表明,M24、M09及M17不同程度地含有黑麦染色体成分,而且电泳谱带差别较大,据此推断,M09为6RL的易位系。因此,生化和分子标记不仅可以用于确定外源片段的存在,而且可以帮助确定染色体组型和外源片段的位置 相似文献
193.
Identification of new mutations in the Cu/Zn superoxide dismutase gene of patients with familial amyotrophic lateral sclerosis. 总被引:6,自引:0,他引:6 下载免费PDF全文
A Pramatarova D A Figlewicz A Krizus F Y Han I Ceballos-Picot A Nicole M Dib V Meininger R H Brown G A Rouleau 《American journal of human genetics》1995,56(3):592-596
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder affecting motor neurons. Although most cases of ALS are sporadic, approximately 10% are inherited as an autosomal dominant trait. Mutations in the Cu/Zn superoxide dismutase gene (SOD 1) are responsible for a fraction of familial ALS (FALS). Screening our FALS kindreds by SSCP, we have identified mutations in 15 families, of which 9 have not been previously reported. Two of the new mutations alter amino acids that have never been implicated in FALS. One of them affects a highly conserved amino acid involved in dimer contact, and the other one affects the active-site loop of the enzyme. These two mutations reduce significantly SOD 1 enzyme activity in lymphoblasts. Our results suggest that SOD 1 mutations are responsible for > or = 13% of FALS cases. 相似文献
194.
Construction and use of a nontoxigenic strain of Pseudomonas aeruginosa for the production of recombinant exotoxin A. 下载免费PDF全文
To express recombinant forms of Pseudomonas aeruginosa exotoxin A in high yield, we have developed a nontoxigenic strain of P. aeruginosa derived from the hypertoxigenic strain PA103. The nontoxigenic strain, designated PA103A, was produced by the excision marker rescue technique to replace the toxA structural gene in PA103 with an insertionally inactivated toxA gene. The PA103A strain (ToxA-) was used subsequently as the host strain for the expression and production of several recombinant versions of exotoxin A, and the results were compared with exotoxin A production in other P. aeruginosa and Escherichia coli strains. Use of the PA103A strain transformed with the high-copy-number pRO1614 plasmid bearing various toxA alleles resulted in final purification yields of exotoxin A averaging 23 mg/liter of culture. By comparison, exotoxin A production in other expression systems and host strains yields approximately 1/4 to 1/10 as much toxin. 相似文献
195.
Le Van Phung Yuki Han Shiro Oka Hisako Hotta Michael D. Smith Prapit Theeparakun Eiko Yabuuchi Ikuya Yano 《FEMS immunology and medical microbiology》1995,12(3-4):259-264
Abstract The serodiagnosis of melioidosis is commonly performed with tests using protein or polysaccharide as antigen. However, due to the low sensitivity, specificity and difficulty in the preparation of the antigens, more simple, precise and reproducible diagnostic tests were required. A purified glycolipid antigen (GL) which is a specific lipid component of Burkholderia pseudomallei has been used in an ELISA. With this antigen, specific immunoglobulin G (IgG) was detected in 49 out of 50 melioidosis sera. IgG was also detected in 2 out of 185 (Japanese) and 16 out of 181 (Vietnamese) control sera. Thus, the sensitivity was 98.0%, and specificity was 98.9% and 91.1% in the Japanese and Vietnamese sera, respectively. When the ELISA and indirect haemagglutination (IHA) tests were combined, a sensitivity of 100% and specificity of 97.8% were achieved. The advantages of the glycolipid antigen are ease of preparation, stability, high sensitivity and specificity. 相似文献
196.
Hong-Bo Qu Xian-En Zhang Wen-Wu Chui Shu-Zheng Zhang Gao-Xiang Li Fan Ouyang 《Biotechnology Techniques》1995,9(6):445-450
Summary A dual-enzyme electrode flow injection system that can simultaneously determine glucose and maltose is used for an on-line study of starch hydrolyses catalysed by amylases. With the working system, determinations can be made every 2 minutes. A 10 L sample size with recycled back-flow minimises any loss of the reaction medium. The production, growth and decay of glucose and maltose concentrations during starch hydrolysis under various enzymatic conditions can thus be closely monitored, making it useful for the study of the catalytic kinetics of amylases and in screening and analysing enzyme systems. 相似文献
197.
Preparation and structural characterization of large heparin-derived oligosaccharides 总被引:4,自引:2,他引:2
Pervin Azra; Gallo Cindy; Jandik Kenneth A; Han Xue-Jun; Linhardt Robert J. 《Glycobiology》1995,5(1):83-95
Porcine mucosal heparin was partially depolymerized with heparinlyase I and then fractionated into low-molecularweight (<5000)and high-molecular-weight (>5000) oligosaccharides by pressurefiltration. The high-molecular-weight oligosaccharide mixture({small tilde}50 wt% of the starting heparin) also containedintact heparin. This intact polymer complicates oligosacsharidepurification. Thus, the low-molecular-weight fraction was usedto prepare homogeneous oligosaccharides for structural characterization.The low-molecular-weight oligosaccharide mixture was first fractionatedby low pressure gel permeation chromatography into size-uniformmixtures of disaccharides, tetrasaccharides, hexasaccharides,octasaccharides, decasaccharides, dodecasaccharides, tetradecasaccharidesand higher oligosaccharides. Each size-fractionated mixturewas then purified on the basis of charge by repetitive semi-preparativestrong-anion-exchange high-performance liquid chromatography.This approach has led to the isolation of 14 homogeneous oligosaccharidesfrom disaccharide to tetradecasaccharide. The purity of theseheparin-derived oligosaccharides was determined by gradientpolyacrylamide gel electrophoresis, analytical strong-anion-exchangehigh-performance liquid chromatography, capillary electrophoresisand one-dimensional nuclear resonance spectroscopy. The structureof these oligosaccharides was established using 600 MHz two-dimensionalnuclear resonance spectroscopy . The spectral methods used includedhomonuclear correlation spectroscopy, nuclear Overhauser effectspectroscopy and heteronuclear multiple quantum coherence spech-clscopy.The 1H/1H connectivities of the protons of each sugar residuein an oligosaccharide were established by two-dimensional homonuclearcorrelation spectroscopy, while 1H/13C assignments were madeusing 1H inverse detection. One- and two-dimensional nuclearresonance spectroscopic analysis of these heparin oligosaccharidesshowed two closely related groups of heparin-oligosaccharidesare afforded by enzymatic depolymerization of heparin. One groupis fully sulphated, having the structures 相似文献
198.
人脑神经元特异性烯醇化酶的纯化方法 总被引:1,自引:0,他引:1
采用改良的Grace层析方法,经一次DEAE-Sephadex A50柱层析即从人脑中纯化了神经元特异性烯醇化酶,比活力为92.1U/mg,纯化倍数为59.4.该酶纯化后,经SDS-聚丙烯酰胺凝胶电泳鉴定为单一蛋白质谱带.此外,还测定了其部分理化性质,其亚单位分子量为45000,等电点pI为4.7,氨基酸组成分析表明其为一种酸性蛋白质;对2-磷酸甘油酸的Km值为5.6×10-4mol/L. 相似文献
199.
200.