酿酒酵母芳樟醇耐受性的工程改造

【背景】芳樟醇具有特殊的香气和多种生物学活性,是食品、医药和化妆品行业的重要原料。随着合成生物学的高速发展,代谢改造微生物进行芳樟醇生物合成是当前研究的一大热点。然而在微生物的生物合成中,芳樟醇对底盘细胞的毒性是一大瓶颈问题,也是其他单萜物质生物合成的共性问题。【目的】建立合理的耐受性改造方法,以提高微生物宿主细胞对芳樟醇的耐受性。【方法】以酿酒酵母BY4741为研究对象,通过对ABC转运蛋白、活性氧调控相关酶及转录调控因子的过表达,考察它们对酿酒酵母芳樟醇耐受性的影响,并通过对酿酒酵母细胞进行定向驯化,筛选耐受性提高的酿酒酵母突变株。【结果】单独过表达ABC转运蛋白(Yor1、Snq2、Pdr5、Pdr15和Pdr18)、ROS调控相关酶(Gre2、Ctt1、Yhb1、Gpx2、Trr1、Trx2和Gsh2)及转录调控因子(Ino2、Yap1、Yap5和Stb5)并不能有效提高酿酒酵母的耐受性,但在传代适应性驯化过程中获得了两株耐受性提高的酿酒酵母突变株,将芳樟醇的致死浓度从430mg/L提高到了645mg/L以上。进一步通过基因组重测序分析揭示了驯化菌株突变位点。其中YBR074W...     

On the origin and colonization of house mice in the Madeira Islands

The skulls and skins of adult house mice from the Madeira Islands have been studied and compared with those from the Salvage Islands and with material from the neighbouring Portuguese mainland man-associated and wild forms, respectively Mus musclus domesticus Rutty, 1772 and M. sprelus Lataste, 1883. Differences between island and mainland populations were found in some of the analysed features. Insular skins of mice were found to be smaller than those of specimens from the mainland. However, in Madeiran and Salvage mice the toothrows were much more developed than in the mainland house mice. It is considered that the causes of these differences lie in the different characteristics of the habitats, mainly food availability, and also in the isolation of populations. Mus musculus domesticus appears to be the only form of the house mouse to have so far successfully colonized the Madeiras.     

First Record of the Peritrich Trichodina diaptomiBasson and Van As, 1991 (Protozoa: Ciliophora) on a South American Calanoid Notodiaptomus deitersi (Poppe, 1890) (Crustacea: Copepoda)

ABSTRACT. The ciliate Trichodina was recorded on the calanoid Notodiaptomus deitersi in a shallow, eutrophic reservoir, located in the centre‐west of Brazil. The species was confirmed as Trichodina diaptomi, a species widely distributed in Europe, Africa, Asia and Australia. It was observed moving freely over the carapace of the copepod, and using its adhesive disc. This is the first record for the species in the Americas.     

Isolation and characterization of 11 microsatellite loci in the mangrove crab, Ucides cordatus (Ocypodidae: Ucidinae)

Eleven polymorphic microsatellite loci were described for the mangrove crab, Ucides cordatus, an important fishery resource on the Brazilian coast. The number of alleles observed at each locus varied between eight and 23. Observed and expected mean heterozygosities were 0.791 and 0.893 respectively. Amplification of all loci was highly successful, under the same polymerase chain reaction conditions. With the exception of P2D3, all loci adhered to the assumptions of the Hardy-Weinberg equilibrium and did not present deviations reflecting linkage disequilibrium. Given this, these markers will be extremely useful in future management programmes for U. cordatus stocks.     

Orally efficacious NR2B-selective NMDA receptor antagonists

A novel series of benzamidines was synthesized and shown to exhibit NR2B-subtype selective NMDA antagonist activity. Compound 31 is orally active in a carrageenan-induced rat hyperalgesia model of pain and shows no motor coordination side effects.     

Discovery of a nonpeptidic small molecule antagonist of the human platelet thrombin receptor (PAR-1)

The synthesis and biological evaluation of a series of nonpeptidic small molecule antagonists of the human platelet thrombin receptor (PAR-1) are described. Optimization of the 5-amino-3-arylisoxazole lead resulted in an approximate 100-fold increase in potency. The most potent of these compounds (54) inhibits platelet activation with IC(50)s of 90 nM against the thrombin receptor agonist peptide (TRAP) and 510 nM against thrombin as the agonist. Further, antagonist 54 fully blocks platelet aggregation stimulated by 1 nM thrombin for 10 min.     

Analysis of maternal effect mutant combinations elucidates regulation and function of the overlap of hunchback and Krüppel gene expression in the Drosophila blastoderm embryo

The metameric organisation of the Drosophila embryo is generated early during development, due to the action of maternal effect and zygotic segmentation and homeotic genes. The gap genes participate in the complex process of pattern formation by providing a link between the maternal and the zygotic gene activities. Under the influence of maternal gene products they become expressed in distinct domains along the anteroposterior axis of the embryo; negative interactions between neighboring gap genes are thought to be involved in establishing the expression domains. The gap gene activities in turn are required for the correct patterning of the pair-rule genes; little is known, however, about the underlying mechanisms. We have monitored the distribution of gap and pair-rule genes in wild-type embryos and in embryos in which the anteroposterior body pattern is greatly simplified due to combinations of maternal effect mutations (staufen exuperantia, vasa exuperantia, vasa exuperantia, bicoid oskar, bicoid oskar torsolike, vasa torso exuperantia). We show that the domains of protein distribution of the gap genes hunchback and Krüppel overlap in wild-type embryos. Based on the analysis of the maternal mutant combinations, we suggest an explanation of how this overlap is generated. Furthermore, our data show that different constellations of gap gene activities provide different input for the pair-rule genes, and thus strongly suggest that the overlap of hunchback and Krüppel in wild-type is functional in the formation of the patterns of pair-rule genes.     

Fertility table and rate of population growth of the predator Supputius cincticeps (Heteroptera: Pentatomidae) on one plant of Eucalyptus cloeziana in the field

The reproductive potential and population growth (r_m and λ) of the predator Supputius cincticeps (Heteroptera: Pentatomidae) were evaluated using a life and fertility table. S. cincticeps was reared on one plant of Eucalyptus cloeziana in the field and fed with Tenebrio molitor (Coleoptera: Tenebrionidae) pupae. Females of this predator had a net reproductive rate (R₀) of 21.02 females/female; an intrinsic rate of population increase (r_m) of 0.041 and finite rate of increase (λ) of 1.042. This resulted in population growth of S. cincticeps in the eucalyptus plant with a doubling time of 17.01 days. This natural enemy can be reared under field conditions with alternative prey for use in biological control. Such individuals of S. cinticeps will be better adapted to field conditions when they are liberated.     

Brood size: a major factor influencing male dimorphism in the non‐pollinating fig wasp Sycobia sp.

• 1 Male polymorphisms have been described in some non‐pollinating fig wasps, as well as in other animals. The proximal basis and the maintenance of alternative male reproductive strategies are either genetic or environmental.
• 2 Here we studied male dimorphism in the non‐pollinating fig wasp Sycobia sp. We conducted experimental manipulations to study the factors influencing offspring male morph allocations and explore a possible basis for the determination and maintenance of male dimorphism in Sycobia sp.
• 3 The results showed that brood size was the major and underlying factor influencing the male morph ratio. When the brood size increases, the wingless male ratio also increases.
• 4 Also, our results indicated that there was no direct maternal control on offspring male morph allocation.
• 5 Male dimorphism in Sycobia sp. probably represents an environmentally determined conditional strategy, which responded to offspring population density at the level of the individual fig.

     

Substrate recognition by the Lyn protein-tyrosine kinase. NMR structure of the immunoreceptor tyrosine-based activation motif signaling region of the B cell antigen receptor

The immunoreceptor tyrosine-based activation motif (ITAM) plays a central role in transmembrane signal transduction in hematopoietic cells by mediating responses leading to proliferation and differentiation. An initial signaling event following activation of the B cell antigen receptor is phosphorylation of the CD79a (Ig-alpha) ITAM by Lyn, a Src family protein-tyrosine kinase. To elucidate the structural basis for recognition between the ITAM substrate and activated Lyn kinase, the structure of an ITAM-derived peptide bound to Lyn was determined using exchange-transferred nuclear Overhauser NMR spectroscopy. The bound substrate structure has an irregular helix-like character. Docking based on the NMR data into the active site of the closely related Lck kinase strongly favors ITAM binding in an orientation similar to binding of cyclic AMP-dependent protein kinase rather than that of insulin receptor tyrosine kinase. The model of the complex provides a rationale for conserved ITAM residues, substrate specificity, and suggests that substrate binds only the active conformation of the Src family tyrosine kinase, unlike the ATP cofactor, which can bind the inactive form.