Phytate dephosphorylation by free and immobilized cells ofSaccharomyces cerevisiae

Summary Saccharomyces cerevisiae in the form of baker's yeast, cells cultivated on a yeast extract-peptone-glucose medium, as well as cells immobilized in 18% (w/v) polyacrylamide gel showed the ability to hydrolyze 1.727 mM sodium phytate solution at 45°C, pH 4.6, in a stirred tank reactor. Seventy percent yield of dephosphorylation was observed after 2 h using a baker's yeast concentration of 5.8 g dry matter per 100 ml. Hydrolytic activity at 1.8–2.0 M Pi min^–1 was observed between 1st and 3rd h of the reaction in cells cultured 24 or 48 h. No inhibition by the substrate was found at sodium phytate concentrations of 0.587–1.727 mM. After 1.5 h of hydrolysis a single, well distinguished peak ofmyo-inositol-triphosphate was the main product found. By means of immobilization the stability of the biocatalyst was enhanced 3.3-fold and reached its half-life at 64 ninety-minute runs.     

大阪鲫鱼（Carassius Auratus cuvieri Temminck et．Schlegel）雌性特异血清蛋白生化特性及免疫细胞化学定位的研究

雌性特异血清蛋白存在于成熟雌性大阪鲫鱼血清中,雄鱼则无。注射雌二醇可诱导雄鱼及成熟雌鱼合成这种蛋白质并引起鱼肝细胞粗面内质网增加,糖元颗粒减少。从诱导的大阪鲫鱼血清中分离出电泳纯的雌性特异血清蛋白的分子量为４６６０００±４０００（ｎ＝１０）,电泳谱带有３条,用纯的雌性特异血海蛋白制备的兔抗鱼抗血清不与雄鱼血清发生免疫反应,而与正常成熟雌鱼及诱导鱼的血清形成１条免疫沉淀线。免疫细胞化学定位诱导及成熟雌鱼的肝细胞核外细胞质有阳性颗粒,在卵母细胞外围有成团的阳性颗粒分布。     

厌氧条件下微量琼脂糖弥散法抑菌试验的建立及应用

作者建立了在厌氧条件下两种微量而敏感的抑菌试验,可用于鉴定蛋白质或多肽类抑菌物质。（１）琼脂糖弥散法：可检测抗菌蛋白抑菌活性,（２）电泳凝胶弥散法：可直接确定存在于ＰＡＧＥ凝胶中抗菌蛋白条带。应用这两种方法,作者首次鉴定出血链球菌培养上清液中存在抑制牙周可疑致病菌的抗菌蛋白。     

Bacterial genome mapping by two-dimensional pulsed-field gel electrophoresis (2D-PFGE)

Macrorestriction mapping is often the first step toward a thorough physical and genetic characterization of a bacterial genome. The problem of deducing the order of partially or completely digested macrorestriction fragments to yield a physical genome map may readily be solved by applying twodimensional pulsed-field gel electrophoresis (2D-PFGE) techniques. These powerful methods are quick and technically easy to perform; specifically, they are independent of DNA probes and should therefore be applicable to any bacterial species irrespective of its prior genetic characterization. In this article, detailed step-by-step protocols are given to set up, run, and evaluate 2D pulsed-field gels. Two basic methods are described: partial/complete 2D gels of one restriction enzyme and complete/complete 2D gels of two different restriction enzymes. Other topics include preparation of bacterial genomic DNA, screening for suitable rare-cutting restriction enzymes and determination of optimal running conditions. Accompanied by many notes, these protocols are meant to offer the novice a sound and rapid access to these important methods.     

Genetic relationships of Brachygobius and related brackish Indo-west Pacific and Australasian genera (Teleostei: Gobiidae)

Horizontal starch gel electrophoresis was employed to analyse genetic variation, at 19 isozyme loci, in six putatively related species of gobies ( Brachygobius aggregatus, B. doriae, Pandaka lidwilli, Pseudogobius olorum, Pseudogobius sp. and Redigobius sp.), using seven other gobiine species as an outgroup. Allozyme data was analysed both cladistically and phenetically, all treatments confirming the monophyly of the group. Pandaka is confirmed as the sister genus to Brachygobius , these genera forming a crown group, with Redigobius and Pseudogobius forming successive sister groups. This scenario is in agreement with morphological hypotheses of phylogeny. However, this lineage appears to be relatively unrelated to other groups within the Gobiinae     

GENETIC VARIABILITY OF GELIDIUM CANARIENSIS (RHODOPHYTA) DETERMINED BY ISOZYME ELECTROPHORESIS1

The populations of Gelidium canariensis (Grunow) Seoane-Camba from the Canary Islands were analyzed for genetic variability by isozyme electrophoresis in 1989 and 1990. Each population was divided into sporophytic and gametophytic subpopulations. Twenty-three to 27 putative alleles corresponding to 22 gene loci were analyzed. Sev-enteen loci were monomorphic in all six subpopulations, and five were polymorphic in at least one subpopulation. Significant deviations from Hardy-Weinberg equilibrium were found. The amount of genetic variability (percentage of polymorphic loci, mean number of alleles per locus, and average gene diversity) of haploid subpopulations was lower than that of diploid subpopulations. No correlation between genetic distance and geographical distance was found. Low genetic differentiation between sporophytic and gametophytic subpopulations of the same locality was obsewed in two populations. The low genetic diversity and genetic differentiation suggest that the genetic structure of the populations of G. canariensis from the Canary Islands is due to a combination of founder effects and the predominance of asexual reproduction. Initial differences in gene frequencies may have persisted because of insufficient time to reach a higher level of differentiation.     

The evolutionary significance of bimodal emergence in the butterfly, Maniola jurtina (Lepidoptera: Satyrinae) (L.).

Most temperate butterflies exhibit a tightly synchronized unimodal adult emergence to facilitate mate location. Exceptions are presumably subject to unusual selection pressure. This study examines the pattern of emergence in Maniola jurtina , which was found to exhibit both unimodal and bimodal emergence patterns at different sites in south-east England. The bimodal pattern was found on chalk grassland; elsewhere the emergence was unimodal. Adults from each emergence peak rarely meet, suggesting that there may be some degree of reproductive isolation. Morphological measurements and electrophoretic analysis of allozyme frequencies are carried out to quantify differentiation between emergence peaks. Captive stock was reared to examine differences in the immature stages. Butterflies from each emergence differ significantly in most morphological variables measured, those from the second peak tending to be smaller. The immature stages differ in morphology and longevity of the egg stage. Allozyme frequencies did not differ between peaks, suggesting that they are not reproductively isolated. Explanations for the maintenance of differences between emergence peaks despite gene flow are discussed. I propose that division of offspring between two emergence times may have evolved to avoid the risk inherent in placing all offspring in one peak which may be rendered inviable by temporal fluctuations in habitat quality.     

Electrophoretic karyotypes of the elm tree pathogen Ophiostoma ulmi (sensu lato)

Pulsed field gel electrophoresis using OFAGE, TAFE, and CHEF systems has been used to more fully characterize karyotypic variation within the two closely related fungal species of Ophiostoma ulmi sensu lato. Twelve wild-type and laboratory strains, representing the less agressive species O. ulmi and both of the biotypes of the more aggressive species O. novo-ulmi were studied and their karyotypes determined. Depending on the strain, a minimum of four to a minimum of eight chromosomal DNA bands were present that fall into three distinct size classes, with one exception. Strain CESSI6K (O. novo-ulmi, North American aggressive subgroup) contains a unique chromosomal DNA band which comigrated near a Saccharomyces cerevisiae chromosome of 0.95 Mb. This unique band was the smallest O. ulmi s. l. chromosomal DNA observed. Seven of the twelve strains shared a common chromosomal DNA banding pattern, whereas each of the other five had a unique karyotype. There was no correlation between chromosome profile and species, as some O. novo-ulmi and O. ulmi strains shared common electrophoretic karyotypes.     

Application of continuous zone electrophoresis to preparative separation of proteins

A comprehensive study of the application of continuous zone electrophoresis to preparative separation of proteins in free solution is presented. First, the influence of electric field strength, buffer residence time in the chamber, sample flow rate, and sample concentration on separation resolution and throughput were studied. Using multiple injections of sample into the electrophoresis chamber, a throughput of 500 mg protein/h was achieved for partially purified model proteins. Experiments on Escherichia coli crude extracts yielded a fivefold purification of beta-galactosidase along with a simultaneous separation of proteins from cell debris in a single step. Experiments correlating the electrophoretic mobility in continuous electrophoresis with the elution behavior in ion-exchange chromatography were performed on more than a dozen proteins which conclusively showed that separation of proteins in continuous zone electrophoresis is governed by net surface charge. Based on these results, the fraction numbers in which the proteins eluted could be correctly predicted. Proteins and enzymes with differences >0.5 M elution molarities in ion-exchange chromatography were separated by continuous zone electrophoresis on a preparative scale (mg/h or g/h) with >90% recovery. This corresponds to a preparative scale separation of proteins and enzymes which differ in apparent electrophoretic mobility by only 0.70 x 10(-5) cm(2)/V . s. (c) 1993 John Wiley & Sons, Inc.     

肺癌患者与肺结核患者血清蛋白的电泳分析

采用SDS聚丙烯酰胺凝胶电泳银染色的方法,对58例肺癌患者和32例肺结核患者的血清蛋白进行比较分析,发现有五种特异蛋白,分别存在于A区、C区、F区和G区。肺癌组和肺结核组比较,存在极显著性差异(P<0.01),提示有早期诊断肺癌的价值,有可能成为临床诊断肺癌的一个重要参考指标。