Fluorometric assay for determining epoxide hydrolase activity

We have developed a rapid screening procedure that enables the screening of hundreds of enzyme samples or variants for epoxide hydrolase activity towards any substrate. The procedure detects the products of the enzymatic reaction via periodate cleavage and remaining fluorescence of carboxyfluorescein.     

Functional analyses of a putative plasma membrane Na+/H+ antiporter gene isolated from salt tolerant Helianthus tuberosus

Jerusalem artichokes (Helianthus tuberosus L.) can tolerate relatively higher salinity, drought and heat stress. In this paper, we report the cloning of a Salt Overly Sensitive 1 (SOS1) gene encoding a plasma membrane Na⁺/H⁺ antiporter from a highly salt-tolerant genotype of H. tuberosus, NY1, named HtSOS1 and characterization of its function in yeast and rice. The amino acid sequence of HtSOS1 showed 83.4 % identity with the previously isolated SOS1 gene from the Chrysanthemum crassum. The mRNA level in the leaves of H. tuberosus was significantly up-regulated by presence of high concentrations of NaCl. Localization analysis using rice protoplast expression showed that the protein encoded by HtSOS1 was located in the plasma membrane. HtSOS1 partially suppressed the salt sensitive phenotypes of a salt sensitive yeast strain. In comparison with wild type (Oryza sativa L., ssp. Japonica. cv. Nipponbare), the transgenic rice expressed with HtSOS1 could exclude more Na⁺ and accumulate more K⁺. Expression of HtSOS1 decreased Na⁺ content much larger in the shoot than in the roots, resulting in more water content in the transgenic rice than WT. These data suggested that HtSOS1 may be useful in transgenic approaches to improving the salinity tolerance of glycophyte.     

AnnexinA7 promotes epithelial–mesenchymal transition by interacting with Sorcin and contributes to aggressiveness in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and metastasis is the major cause of the high mortality of HCC. In this study, we identified that AnnexinA7 (ANXA7) and Sorcin (SRI) are overexpressed and interacting proteins in HCC tissues and cells. In vitro functional investigations revealed that the interaction between ANXA7 and SRI regulated epithelial–mesenchymal transition (EMT), and then affected migration, invasion, and proliferation in HCC cells. Furthermore overexpression/knockdown of ANXA7 was remarkably effective in promoting/inhibiting tumorigenicity and EMT in vivo. Altogether, our study unveiled a mechanism that ANXA7 promotes EMT by interacting with SRI and further contributes to the aggressiveness in HCC, which provides a novel potential therapeutic target for preventing recurrence and metastasis in HCC.Subject terms: Medical research, Genetics research     

树叶凋落物在受酸性矿山废水污染溪流中的分解

为了解华南地区酸性矿山废水对溪流中树叶分解的影响,在广东省大宝山矿区附近的1条受酸性矿山废水污染（pH值为2．7—3．4且富含多种重金属元素）的3级溪流中,利用2种孔径（5ram的网袋和0．1ram的布袋）的分解网袋对2种树叶（人面子和蒲桃）进行了为期101d的树叶分解研究。结果表明,人面子树叶网袋和布袋中的树叶干重剩余率分别为39％和48％,而蒲桃树叶网袋和布袋中的干重剩余率仍保持较高的水平,分别为61％和70％。根据指数衰减模型计算出树叶分解的半衰期,人面子树叶在网袋和布袋中的分解半衰期分别为57d和69d,而蒲桃树叶则分别为14-4d和217d。蒲桃树叶的分解速率明显比人面子树叶慢。在网袋中定殖的底栖动物主要是集食者,其中优势类群为摇蚊幼虫,占底栖动物个体总数的99％。摇蚊种群在网袋中的数量波动对2种树叶分解速率的影响并不明显。结果表明,受酸性矿山废水的影响,底栖动物群落的多样性大为减少。同时由于各种金属氧化物在树叶表面的不断沉淀,使树叶处于缺氧状态,抑制了微生物的活动,导致树叶分解速率大为降低。     

Flow cytometry-assisted rapid isolation of recombinant Plasmodium berghei parasites exemplified by functional analysis of aquaglyceroporin

The most critical bottleneck in the generation of recombinant Plasmodium berghei parasites is the mandatory in vivo cloning step following successful genetic manipulation. This study describes a new technique for rapid selection of recombinant P. berghei parasites. The method is based on flow cytometry to isolate isogenic parasite lines and represents a major advance for the field, in that it will speed the generation of recombinant parasites as well as cut down on animal use significantly. High expression of GFP during blood infection, a prerequisite for robust separation of transgenic lines by flow cytometry, was achieved. Isogenic recombinant parasite populations were isolated even in the presence of a 100-fold excess of wild-type (WT) parasites. Aquaglyceroporin (AQP) loss-of-function mutants and parasites expressing a tagged AQP were generated to validate this approach. aqp^? parasites grow normally within the WT phenotypic range during blood infection of NMRI mice. Similarly, colonization of the insect vector and establishment of an infection after mosquito transmission were unaffected, indicating that AQP is dispensable for life cycle progression in vivo under physiological conditions, refuting its use as a suitable drug target. Tagged AQP localized to perinuclear structures and not the parasite plasma membrane. We suggest that flow-cytometric isolation of isogenic parasites overcomes the major roadblock towards a genome-scale repository of mutant and transgenic malaria parasite lines.     

广东金山温泉沉积物中原核与真核微生物多样性初步分析

[目的]本研究旨在采用不同的PCR引物对广东省恩平市金山温泉的高温水底沉积物微生物多样性进行初步的分析.[方法]采用改进的玻璃珠法抽提温泉沉积物中环境基因组DNA,通过对用4对引物分别扩增得到的原核微生物16S rRNA基因和真核微生物ITS序列的分析,将所得到的数据与国际基因数据库GenBank进行相似性比较并构建系统发育树.[结果]研究发现原核类群G的 14个优势克隆中7个都属于蛭弧菌属(Bdellovibrio).与它们最相似的序列是从海洋中分离到的两个菌株 Bacteriovorax sp. NE1 (EF092445)和Bdellovibrio sp. JS5 (AF084859),相似性分别为96%和99%.原核类群X的4个序列主要属于蓝细菌类群,其中JS-X2与在美国黄石公园温泉发现的Uncultured Cyanobacterium (L35331)有95%的相似性,并且与已经全基因组测序的嗜热蓝细菌聚球藻Thermosynechococcus elongatus BP-1 (47118315)有89%的相似性.真核类群Z有三个类群,分别是Penicillium sp.,Lodderomyces sp.和Gloeotinia sp..其中大部分序列与青霉属相似性在88%～ 90%之间.[结论]所得到的结果显示金山温泉中的微生物多样性十分丰富.     

银额果蝇的B染色体研究 2.昆明群体的生活力和B染色体的关系

本文分析了钮额果蝇的生活力和B染色体的关系。结果表明,银额果蝇的B染色体在其生长和发育过程中具双重性调节作用。一方面,B染色体以单一的形式存在于单雌系核型中,即1B或2Bs,可刺激生长、发育,增强生活力。另一方面,在单雌系的细胞中存在多条B染色体,并形成核型多态性,这似乎对生长和发育又有一定的抑制作用而减弱其生活力。     

Isolation and culture of daylily mesophyll protoplasts

Mesophyll protoplasts were isolated from leaf tissues of a diploid daylily (HemerocallisxRed Magic) by enzymatic digestion with a solution containing 0.5% Pectolyase Y-23, 0.1% Cellulase R-10, 0.1% Driselase, 0.6 M sorbitol and half-strength MS inorganic salts. When cultured on MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA, the protoplasts underwent sustained division to produce multicellular colonies. The optimal plating density for cell division was 0.5 × 10⁵ protoplasts/ml. The highest plating efficiency was obtained in cultures grown in media solidified with 0.2% Gelrite. Under these conditions, formation of colonies occurred from 14% of cultured protoplasts. Calli were recovered from 9 colonies only after the cultures were treated with a conditioned medium. Intact plants were regenerated from protoplast-derived calli through organogenesis.Abbreviations BA 6-benzylaminopurine - FDA fluorescein diacetate - GA₃ gibberellic acid - MS medium Murashige and Skoog (1962) medium - NAA 1-naphthaleneacetic acid     

Cleavage of territrems by alkaline hydrogen peroxide: Isolation and characterization of the aromatic moiety of territrems

The chemical reaction of cleavaging territrem B to give 3,4,5-trimethoxy benzoic acid by alkaline hydrogen peroxide was investigated. The method was applied for confirmation of the chemical structure of the aromatic moiety of territrem A, A’, B, and B’. The physicochemical properties of the aromatic cleavage product of territrem Aindicated the structure as 3,4-methylendioxy, 5-methoxy benzoic acid (or 4-methoxy, 6-carboxy, 1, 3-benzodioxole). The experiment also gave the evidences that territrem A and A’, on the other hand territrem B and B’ have the identical aromatic moieties on their structures.     

Phylogenetics of early branching eudicots: Comparing phylogenetic signal across plastid introns, spacers, and genes

Recent phylogenetic analyses revealed a grade with Ranunculales,Sabiales,Proteales,Trochodendrales,and Buxales as first branching eudicots,with the respective positions of Proteales and Sabiales still ...