NAD-linked aldehyde dehydrogenase for aerobic utilization of L-fucose and L-rhamnose by Escherichia coli.

Mutant analysis revealed that complete utilization of L-fucose and L-rhamnose by Escherichia coli requires the activity of a common NAD-linked aldehyde dehydrogenase which converts L-lactaldehyde to L-lactate. Mutations affecting this activity mapped to the ald locus at min 31, well apart from the fuc genes (min 60) encoding the trunk pathway for L-fucose dissimilation (as well as L-1,2-propanediol oxidoreductase) and the rha genes (min 88) encoding the trunk pathway for L-rhamnose dissimilation. Mutants that grow on L-1,2-propanediol as a carbon and energy source also depend on the ald gene product for the conversion of L-lactaldehyde to L-lactate.     

鲫鱼尾部神经分泌系统显微和亚显微结构的季节性变化

鲫鱼尾部神经分泌系统的神经分泌细胞和它的轴突中可观察到各种不同电子密度的颗粒。在性腺各个不同的发育阶段,该系统的分泌物具有累积、充满、释放和恢复这样一种周期性变化,由此说明鲫鱼的尾部神经分泌系统和它的生殖有关。     

An immunocytochemical method for the localization of fibronectin in Araldite-embedded specimens

Summary The detection of fibronectin (FN) in osmium-fixed and Araldite-embedded frog skin fragments was studied using a modification of Baskin's procedure (Baskin et al. 1979). Following the removal of Araldite from the semi-thin sections (0.5–1.0 m) with ethanol-NaOH solution, the sections were bleached with hydrogen peroxide. FN was detected by indirect immunoperoxidase method. For precise localization of FN, careful attention was paid to the temperature, antibody concentrations and the quality of the ethanol-NaOH solution. Our results were in agreement with those that we had obtained previously for polyethylene glycol (PEG) sections, suggesting that the present procedure is useful for the detection of FN in Araldite-embedded biological specimens.     

Sequence homology in the metalloproteins; purple acid phosphatase from beef spleen and uteroferrin from porcine uterus

The primary structures of purple acid phosphatase and uteroferrin, two iron-binding glycoproteins isolated from beef spleen and porcine uterine fluids, respectively, have been examined by a combination of tandem mass spectrometry and classical Edman sequencing methods. Reported here are amino acid sequence data covering more than 90% of the primary structures for these two proteins. The sequence data reveal an unexpectedly high degree of homology, greater than 90%, for these two proteins.     

水翁花蕾和水翁叶精油的化学成分研究

水翁的花营和鲜叶经水蒸汽蒸馏得到一种淡黄色的精油,前者出油率为0.18％。后者为0.08％。我们应用毛细管气相色谱,气相色谱/质谱联用,红外光谱和紫外光谱等方法,对两种精油进行化学分成分析,分别鉴定出35个和27个已知化学成分。两者相同的化学成分有:β-罗勒烯(Z)、β-罗勒烯(E)、α-蒎烯、β-蒎烯、月桂烯、小茴香烯、香叶醇、顺式-丁香烯、橙花叔醇等23个化学成分,分别占花营精油全油的90％和叶精油95％以上。     

Influence of sucrose concentration and phosphate limitation on citric acid production by immobilized cells of Aspergillus niger

Summary Immobilized cells of Aspergillus niger needed a lower initial sucrose concentration than free cells in order to obtain maximal yields of citric acid production. High sucrose concentrations led to reduced yields and increased polyol formation (glycerol, erythritol, arabitol). Continuous fermentation with media containing low sugar concentrations prevented the formation of polyols. The change from nitrogen-limited to phosphate-limited precultivation of immobilized spores significantly increased the productivity of the mycelium. The ratio of citric acid to residual sugar in the effluent distinctly lay in the direction of citric acid. Inside the alginate beads mainly large bulbous cells were observed.     

A method of isolation of apical membranous sheets from frog urinary bladder epithelium by stripping with gelatin

We have developed a technique for recovering apical membranous sheets from amphibian urinary bladders by gelatin stripping. The tissue is mounted on a lucite support and the apical surface is first stuck onto a gelatin-coated glass slide at 30 degrees C. This sandwich is then chilled on ice and the bladder is pulled away from the slide. Preliminary results indicate that this simple technique could be used to remove membranous apical sheets of various sizes, almost devoid of cytoplasmic contamination and without significant damage to the underlying cell structures. The method could also be adapted to prepare perforated cells and to study the cohesive forces between the different layers of the tissue.     

Oligodeoxynucleotide-directed cleavage and repair of a single-stranded vector: a method of site-specific mutagenesis

A simple and efficient site-specific mutagenesis method is described. First, a single-stranded (ss) circular vector is linearized at the site where the desired mutation will be introduced. To do this, an oligodeoxynucleotide complementary to the target region of the ss vector and containing a restriction enzyme recognition sequence is annealed to the circular ss vector, and the partial double-strand formed is subsequently cleaved with that enzyme. Then, another oligodeoxynucleotide spanning the nick and carrying the mutation is annealed to the linearized ss DNA template and the annealed mixture is used directly to transform Escherichia coli without prior enzymatic DNA synthesis in vitro. The procedure has been applied successfully to constructing insertion, deletion, and point mutations in both M13 phage vectors and plasmid vectors containing the f1 origin of replication.     

Distribution of lipid-binding regions in human apolipoprotein B-100

The distribution of lipid-binding regions of human apolipoprotein B-100 has been investigated by recombining proteolytic fragments of B-100 with lipids and characterizing the lipid-bound fragments by peptide mapping, amino acid sequencing, and immunoblotting. Fragments of B-100 were generated by digestion of low-density lipoproteins (LDL) in the presence of sodium decyl sulfate with either Staphylococcus aureus V8 protease, pancreatic elastase, or chymotrypsin. Particles with electron microscopic appearance of native lipoproteins formed spontaneously when detergent was removed by dialysis from enzyme digests containing fragments of B-100 and endogenous lipids, or from incubation mixtures of delipidated B-100 fragments mixed with microemulsions of exogenous lipids (cholesteryl oleate and egg phosphatidylcholine). Fractionation of the recombinant particles by isopycnic or density gradient ultracentrifugation yielded complexes similar to native LDL with respect to shape, diameter, electrophoretic mobility, and surface and core compositions. Circular dichroic spectra of these particles showed helicity similar to LDL but a somewhat decreased content of beta-structure. Most of the fragments of B-100 were capable of binding to lipids; 12 were identified by direct sequence analysis and 14 by reaction with antisera against specific sequences within B-100. Our results indicate that lipid-binding regions of B-100 are widely distributed within the protein molecule and that proteolytic fragments derived from B-100 can reassociate in vitro with lipids to form LDL-like particles.