Genetic characterization of <Emphasis Type="Italic">psp</Emphasis> encoding the DING protein in <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> SBW25

Background  

DING proteins constitute a conserved and broadly distributed set of proteins found in bacteria, fungi, plants and animals (including humans). Characterization of DING proteins from animal and plant tissues indicated ligand-binding ability suggesting a role for DING proteins in cell signaling and biomineralization. Surprisingly, the genes encoding DING proteins in eukaryotes have not been identified in the eukaryotic genome or EST databases. Recent discovery of a DING homologue (named Psp here) in the genome of Pseudomonas fluorescens SBW25 provided a unique opportunity to investigate the physiological roles of DING proteins. P. fluorescens SBW25 is a model bacterium that can efficiently colonize plant surfaces and enhance plant health. In this report we genetically characterize Psp with a focus on conditions under which psp is expressed and the protein exported.     

Altering coenzyme specificity of <Emphasis Type="Italic">Pichia stipitis</Emphasis> xylose reductase by the semi-rational approach CASTing

Background  

The NAD(P)H-dependent Pichia stipitis xylose reductase (PsXR) is one of the key enzymes for xylose fermentation, and has been cloned into the commonly used ethanol-producing yeast Saccharomyces cerevisiae. In order to eliminate the redox imbalance resulting from the preference of this enzyme toward NADPH, efforts have been made to alter the coenzyme specificity of PsXR by site-directed mutagenesis, with limited success. Given the industrial importance of PsXR, it is of interest to investigate further ways to create mutants of PsXR that prefers NADH rather than NADPH, by the alternative directed evolution approach.     

Nanobiotechnology: quantum dots in bioimaging

Many biological systems, including protein complexes, are natural nanostructures. To better understand these structures and to monitor them in real time, it is becoming increasingly important to develop nanometer-scale signaling markers. Single-molecule methods will play a major role in elucidating the role of all proteins and their mutual interactions in a given organism. Fluorescent semiconductor nanocrystals, known as quantum dots, have several advantages of optical and chemical features over the traditional fluorescent labels. These features make them desirable for long-term stability and simultaneous detection of multiple signals. Here, we review current approaches to developing a biological application for quantum dots.     

A novel butyrolactone derivative inhibited apoptosis and depressed integrin beta4 expression in vascular endothelial cells

To understand the effects of a novel butyrolactone derivative, 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3H)-one (3BDO), on the apoptosis of vascular endothelial cells (VECs), we exposed 3BDO (20-60 microg/ml) to VECs deprived of serum and FGF-2 for 24 and 48 h, respectively. The results showed that 3BDO (20-60 microg/ml) increased VEC viability and inhibited VEC apoptosis induced by deprivation of serum and FGF-2 in a very weak dose-dependent manner. During this process, integrin beta4 expression was depressed, but the level of reactive oxygen species (ROS) was not changed. The data suggested that 3BDO (20-60 microg/ml) could inhibit VEC apoptosis and suppress integrin beta4 expression, but it could not depress the ROS level induced by deprivation of serum and FGF-2.     

Genetic diversity and biogeography of the traditional Chinese medicine, Gardenia jasminoides, based on AFLP markers

Gardenia jasminoides Ellis is used in traditional Chinese medicine (TCM) in China. Levels of genetic variation and patterns of population structure within and among eight wild or cultivated populations of G. jasminoides Ellis in China were investigated using amplified fragment length polymorphism (AFLP) markers. Of the 11 primers screened, four produced highly reproducible AFLP bands. Using these primers, 244 discernible DNA fragments were generated with 165 bands (67.6%), were polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 36.89% to 59.43%. Genetic diversity within populations ranged from 0.2086 to 0.3108, averaging 0.2392 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (76.59%), Shannon's index analysis (64.8%) and AMOVA analysis (72.75%). No significant statistical differences (analysis of molecular variance [AMOVA], p = 0.0639) in AFLP variation were found between regions. However, the variance among populations and within populations differed significantly (p < 0.001). An indirect estimate of historical levels of gene flow (N_m = 1.7448) was consistent with the high mean genetic identity (mean I = 0.9263) found among populations. There is an association between geographic and genetic distances between populations. Presently gene change exists between populations.     

Genomic growth hormone gene polymorphisms in native Chinese chickens

Chicken growth hormone (cGH), a polypeptide hormone synthesized in and secreted by the pituitary gland, is involved in a wide variety of physiological functions such as growth, body composition, egg production, aging, and reproduction. Chicken growth hormone polymorphisms have been reported to be associated with certain phenotypes. Our objective is to investigate the GH gene polymorphism in selected strains of native Chinese chickens. Yellow Wai Chow GH gene was characterized by sequencing and was found to have one silent substitution, 31 insertions, and other substitutions spread among the introns. In addition, a novel Mspl site has been identified and characterized in the first intron. Allele frequencies of the intron 1 polymorphism were characterized among 28 populations of native Chinese chickens. Thus, polymorphism of the cGH gene may be useful in phylogenetic analysis, as well as in the design of breeding programs.     

椴树阔叶红松林群落主要树种径级结构研究

椴树阔叶红松林群落主要树种径级结构研究孙伟中代力民章一平（中国科学院沈阳应用生态研究所,１１００１５）ＳｔｕｄｙｏｆＤｉａｍｅｔｅｒＢｒｅａｓｔＨｉｇｈＳｔｒｕｃｔｕｒｅｏｆｔｈｅＭａｉｎＴｒｅｅｓｉｎＴｉｌｉａＢｒｏａｄｌｅａｖｅｄＫｏｒｅａｎＰ...     

新型有原始肢突胴甲鱼的发现及胴甲鱼早期演化的初步探讨

本文记述的曲靖始突鱼(Procondylolepis qujingensis gen.et sp.nov.)是近几年在云南曲靖早泥盆世地层中发现的有肢突胴甲鱼一原始类型。它和已知胴甲鱼(包括早泥盆世无肢突的和中、晚泥盆世有“盔”状肢突的)不同的最大特点是在其肩带与胸鳍相接的肩关节处有原始的肢突和简单的关节窝;胸鳍甲近端的关节区很小。它展现出胴甲鱼类这一高度特化、长期使人迷惑不解的肢突,在胴甲鱼演化史上发展变化的梗概,填补了肢突从无到有中间的缺环,使人了解到胸鳍的具体结构。文中主要根据肢突的有无和特化程度等,对胴甲鱼早期演化史作了初步探讨,将胴甲鱼分为无肢突超目(Abrachicondylia)和有肢突超目(Brachicondylia)两大部分。始突鱼则代表有肢突超目一早期成员。     

Osmomechanical stress selectively regulates translocation of protein kinase C isoforms

We have investigated the contribution of lipid rafts to activation of the NADPH oxidase enzyme system in neutrophils. Membrane-bound NADPH oxidase subunits are present in the lipid raft compartment of neutrophils. Cytosolic NADPH oxidase components are mainly absent from but are recruited to rafts upon Fcγ receptor activation. In parallel, protein kinase C isotypes are recruited to the rafts. Kinetic analysis of NADPH oxidase activation revealed that rafts determine the onset but not the maximal rate of enzyme activity. Thus lipid rafts serve to physically juxtapose the NADPH oxidase effector, protein kinase C and Fcγ receptor, resulting in efficient coupling.     

Angiostatin production in cultivation of recombinant Pichia pastoris fed with mixed carbon sources

A recombinant strain of Pichia pastoris with a phenotype of Mut^S was used to produce angiostatin. Due to the low methanol consumption rate of this strain, both methanol and glycerol feedings, that produced oscillation in dissolved O₂ concentration, were used during the expression phase to improve cell growth and angiostatin expression. However, enhanced cell growth led to nitrogen limitation that suppressed further production of angiostatin, but addition of ammonia allowed angiostatin concentration to reach 108 mg l^–1 after an expression period of 96 h. The ratio of consumed glycerol to methanol of 1.5:1 (w/w) in the expression phase suggested that methanol played an important role in the metabolism of carbon sources.