枯草芽孢杆菌SC2-4-1葡聚糖酶基因gluB的克隆及序列分析

β-1,3-1,4-葡聚糖酶活性检测结果表明,从辣椒根际筛选的拮抗菌枯草芽孢杆菌(Bacillus subtilis)SC2-4-1能产生β-1,3-1,4-葡聚糖酶.以菌株SC2-4-1的基因组DNA为模板,用PCR方法克隆了该菌的葡聚糖酶基因gluB,其开放阅读框为711bp,编码237个氨基酸.Blast分析,该序列与已报道的多粘类芽孢杆菌(Paenibacillus polymyxa)ATCC 842的β-1,3-1,4-葡聚糖酶基因gluB相似性为85%.所得基因序列的系统发育分析显示,该基因属于β-1,3-1,4-葡聚糖酶基因.DNAMAN软件比对,所得葡聚糖酶氨基酸序列具有催化裂解β-1,3-和β-1,4-糖苷键的葡聚糖酶活性位点.     

寡糖对向日葵叶片细胞病程相关蛋白及细胞壁物质的诱导作用

用寡糖对向日葵叶片进行喷雾,研究了该寡糖对向日葵叶片细胞内几丁质酶和β-1,3-葡聚糖酶2种病程相关蛋白(PRs蛋白)以及木质素、富含羟脯氨酸蛋白(HRGP)2种细胞壁物质含量的影响.结果表明,经寡糖处理后,向日葵叶片中β-1,3-葡聚糖酶和几丁质酶的活性均升高,最高值分别比同期CK增加36.38%和6.35%;木质素及HRGP含量也诱导增加,最高值分别比同期CK显著增加39.15%和47.13%.在寡糖处理后接种病原菌的向日葵叶片中,β-1,3-葡聚糖酶及细胞壁物质被诱导合成,且合成量均较单独寡糖处理与单独接种锈菌处理要高.研究发现,诱导向日葵幼苗叶片的几丁质酶、β-1,3-葡聚糖酶活性增强和细胞壁木质素、HRGP含量增加,可能是寡糖预处理增强向日葵抗锈性的内在机制.     

β-1,3-1,4葡聚糖酶基因克隆表达及其抗菌活性与机理研究进展

β-1,3．1,4-葡聚糖酶是一类能水解β-1,3．糖苷键和β-1,4-糖苷键的酶,因其主要分解大麦中的β-1,3-1,4-葡聚糖和细菌地衣多糖,所以又称地衣多糖酶。综述了β-1,3．1,4-葡聚糖酶基因的克隆表达及其抗菌活性与机理最新研究进展。     

细菌编码β-1,3-1,4-葡聚糖酶催化活性优化方法

β-1,3-1,4-葡聚糖酶是一类专一降解β-葡聚糖的内切水解酶。高效β-葡聚糖酶在啤酒酿造工业上具有十分重要的应用价值。目前,研究较多的β-1,3-1,4-葡聚糖酶主要来源于细菌。文中概述了细菌编码β-1,3-1,4-葡聚糖酶的分子生物学性质,并且从蛋白分子改造、表达调控和发酵条件优化三方面阐述了其催化活性提高的方法和成果。     

细菌β-1,3-1,4-葡聚糖酶分子改造和表达策略

β-1,3-1,4-葡聚糖酶作为一种饲料和食品添加剂有着广泛用途。迄今在杆菌、梭菌、瘤胃细菌、真菌、高等植物中都发现了β-1,3-1,4-葡聚糖酶。综述了细菌来源β-1,3-1,4-葡聚糖酶的性质、结构、分子改造与表达研究进展。     

淀粉液化芽孢杆菌β1-1,3-1,4-葡聚糖酶基因的克隆及表达

为了比较不同的表达系统对β-1,3-1,4-葡聚糖酶基因(bgl)的效果,本研究将高产β-1,3-1,4-葡聚糖酶的淀粉液化芽孢杆菌Bacillus amyloliquefaciens BS5582的bgl基因(GenBank Accession No.EU623974)克隆到3种不同的质粒载体中,即构建pEGX-4T-1-bgl、pET20b(+)-bgl和pET28a(+)-bgl重组质粒.比较了pEGX-4T-1-bgl,在不同Escherichia coli宿主中表达效果,以及pET20b(+)-bgl和pET28a(+)-bgl在E coli BL21(DE3)中的表达效果.结果表明,E. coli BL21(DE3)-pET28a(+)-bgl能够表达最高的重组β-1,3-1,4-葡聚糖酶酶活,其总酶活可达(322.0±8.8)U/mL,是出发菌在最适摇瓶发酵条件下产酶活的40.1%.对该重组菌的产酶条件进行了分析,结合IPTG和乳糖协同的诱导作用,在基础产酶培养基中产最高总酶活为(1883.3±45.8)U/mL,表明其具有良好的工业应用价值.     

淀粉液化芽孢杆菌b-1,3-1,4-葡聚糖酶基因的克隆及表达的优化

为了比较不同的表达系统对β-1,3-1,4-葡聚糖酶基因(bgl)的效果,本研究将高产β-1,3-1,4-葡聚糖酶的淀粉液化芽孢杆菌Bacillus amylolique faciens BS5582的bgl基因(GenBank Accession No.EU623974)克隆到3种不同的质粒载体中,即构建pEGX-4T-1-bgl、pET20b(+)-bgl和pET28a(+)-bgl重组质粒。比较了pEGX-4T-1-bgl在不同Escherichiacoli宿主中表达效果,以及pET20b(+)-bgl和pET28a(+)-bgl在E.coliBL21(DE3)中的表达效果。结果表明,E.coliBL21(DE3)-pET28a(+)-bgl能够表达最高的重组β-1,3-1,4-葡聚糖酶酶活,其总酶活可达(322.0±8.8)U/mL,是出发菌在最适摇瓶发酵条件下产酶活的40.1%。对该重组菌的产酶条件进行了分析,结合IPTG和乳糖协同的诱导作用,在基础产酶培养基中产最高总酶活为(1883.3±45.8)U/mL,表明其具有良好的工业应用价值。     

枯草芽孢杆菌产β-1,3-1,4-葡聚糖酶的响应面优化

【目的】采用响应面法(RSM)优化枯草芽孢杆菌5 L发酵罐产β-1,3-1,4-葡聚糖酶的发酵条件。【方法】利用Box-Behnken设计和方差分析。【结果】获得最佳发酵条件为:转速、通气量和培养基pH分别为500 r/min、1.05 vvm和5.08,发酵时间仅为22 h产β-1,3-1,4-葡聚糖酶活力达2 294.4 U/mL。【结论】实验结果表明响应面法优化5 L发酵罐发酵产β-1,3-1,4-葡聚糖酶的条件合理可行。     

解淀粉芽孢杆菌β-1,3-1,4-葡聚糖酶的高效表达

目的:克隆解淀粉芽孢杆菌β-1,3-1,4-葡聚糖酶基因(bglA)使其在解淀粉芽孢杆菌CICIM B4081中高效表达,并对重组酶进行酶学性质研究.方法:以解淀粉芽孢杆菌(CICIM B4801)染色体DNA为模板,经过PCR扩增得到了大小约为0.8kb的β-1,3-1,4-葡聚糖酶基因(bglA),构建了重组表达质粒pQ-bglA,通过电转化的方法将其转化人解淀粉芽孢杆菌(CICIM B4801)中.结果:得到了能高效表达β-1,3-1,4-葡聚糖酶的重组解淀粉芽孢杆菌.在250mL摇瓶条件下,重组菌分解地衣多糖的胞外最高酶活达到了1515.7U/mL,重组酶的最适作用温度为55℃,最适反应pH值为6.5.结论:重组菌的β-1,3-1,4-葡聚糖酶的酶活为原始菌株的11.84倍,实现了bglA基因在解淀粉芽孢杆菌中的高效表达.     

微生物1,3-1,4-β-葡聚糖酶蛋白质改造及工业应用研究进展

1,3-1,4-β-葡聚糖酶(E.C.3.2.1.73)是一种重要的工业用酶,其可以通过特异性切割毗邻β-1,3-糖苷键的β-1,4-糖苷键将β-葡聚糖或地衣多糖降解为纤维三糖和纤维四糖。微生物β-葡聚糖酶属于糖苷水解酶家族16,其三维结构为卷心蛋糕状的逆向β-片层结构。文中综述了近些年来β-葡聚糖酶在工业上的应用情况及酶蛋白质工程改造的研究进展,并对其研究前景进行了展望。     

Antagonistic Coevolution and Sex

One of the leading hypotheses for the maintenance of sexual reproduction is the Red Queen hypothesis. The underlying premise of the Red Queen hypothesis is that parasites rapidly evolve to infect common host genotypes. This response by parasites could result in the long-term maintenance of genetic variation and may favor sexual reproduction over asexual reproduction. The underlying ideas present a wonderful microcosm for teaching evolution. Here I present the reasons for why sex is anomalous for evolutionary theory, the rationale underlying the Red Queen hypothesis, and some empirical studies of the Red Queen hypothesis using a freshwater snail. The empirical results are consistent with the Red Queen hypothesis. In addition, the distribution of sexual and asexual reproduction in the snail leads naturally to thinking about coevolution in a geographic mosaic of parasite-mediated natural selection.     

拮抗Bacillus subtilis的筛选及发酵条件对拮抗能力的影响

筛选了3株对大肠埃希菌具有拮抗活性的枯草芽胞杆菌,并对这3个菌株的拮抗能力最强的Bf菌株的发酵条件进行了初步研究。先将不同枯草芽胞杆菌菌株与株大肠埃希菌进行拮抗试验,根据抑菌率,筛选出对大肠埃希菌拮抗效果较好的枯草芽胞杆菌Bf。然后,在不同pH值、温度以及不同的培养时间下观察记录Bf对大肠埃希菌的拮抗作用。根据实验结果分析得到:筛选到的枯草芽胞杆菌在pH值为7.0,37℃下培养40 h后抑菌效果最好。     

内生拮抗放线菌

正植物内生菌是一种潜在的重要微生物资源,它能促进宿主植物生长、增强植物抗逆性及增强其抗病虫害能力等,具有开发成植物生长促进剂和生物农药的巨大潜力[1-2]。相对于植物内生真菌和其他内生细菌的广泛深入研究,有关植物内生放线菌生物学作用及其活性物质的研究相对滞后,仍然有许多领域值得进一步深入研究。随着各生物学领域的不断发展,近年来国内相关研究较活跃[3-6],植物内生放线菌更多独特的生物学特性将会被发现,也将作为新兴的     

Antagonistic Parent-Offspring Co-Adaptation

Background

In species across taxa, offspring have means to influence parental investment (PI). PI thus evolves as an interacting phenotype and indirect genetic effects may strongly affect the co-evolutionary dynamics of offspring and parental behaviors. Evolutionary theory focused on explaining how exaggerated offspring solicitation can be understood as resolution of parent-offspring conflict, but the evolutionary origin and diversification of different forms of family interactions remains unclear.

Methodology/Principal Findings

In contrast to previous theory that largely uses a static approach to predict how “offspring individuals” and “parental individuals” should interact given conflict over PI, we present a dynamic theoretical framework of antagonistic selection on the PI individuals obtain/take as offspring and the PI they provide as parents to maximize individual lifetime reproductive success; we analyze a deterministic and a stochastic version of this dynamic framework. We show that a zone for equivalent co-adaptation outcomes exists in which stable levels of PI can evolve and be maintained despite fast strategy transitions and ongoing co-evolutionary dynamics. Under antagonistic co-adaptation, cost-free solicitation can evolve as an adaptation to emerging preferences in parents.

Conclusions/Significance

We show that antagonistic selection across the offspring and parental life-stage of individuals favors co-adapted offspring and parental behavior within a zone of equivalent outcomes. This antagonistic parent-offspring co-adaptation does not require solicitation to be costly, allows for rapid divergence and evolutionary novelty and potentially explains the origin and diversification of the observed provisioning forms in family life.     

拮抗菌抗菌谱及发酵液拮抗能力测定的新方法

目的:确定测定拮抗细菌枯草芽孢杆菌A的抗菌谱及发酵液拮抗能力的方法。方法:平板对峙法测定抗菌谱,三明治法、管碟法、滤纸片法和双层打孔法测定发酵液的拮抗能力。结果:用点接拮抗细菌单菌落的平板对峙法测得菌株A对玉米小斑病菌等10种病原真菌均有拮抗作用;用双层打孔法测定菌株A发酵液拮抗能力,发现在培养条件为温度32℃,溶氧量为300mL三角瓶培养基装量为30mL,接种量5%,初始pH值7.5时发酵液拮抗能力最强。结论:点接拮抗细菌单菌落的平板对峙法适合用来测定菌株A的抗菌谱,双层打孔法适合用来测定菌株A发酵液的拮抗能力。     

Antagonistic and Synergistic Interactions Among Predators

The structure and dynamics of food webs are largely dependent upon interactions among consumers and their resources. However, interspecific interactions such as intraguild predation and interference competition can also play a significant role in the stability of communities. The role of antagonistic/synergistic interactions among predators has been largely ignored in food web theory. These mechanisms influence predation rates, which is one of the key factors regulating food web structure and dynamics, thus ignoring them can potentially limit understanding of food webs. Using nonlinear models, it is shown that critical aspects of multiple predator food web dynamics are antagonistic/synergistic interactions among predators. The influence of antagonistic/synergistic interactions on coexistence of predators depended largely upon the parameter set used and the degree of feeding niche differentiation. In all cases when there was no effect of antagonism or synergism (a _ij =1.00), the predators coexisted. Using the stable parameter set, coexistence occurred across the range of antagonism/synergism used. However, using the chaotic parameter strong antagonism resulted in the extinction of one or both species, while strong synergism tended to coexistence. Whereas using the limit cycle parameter set, coexistence was strongly dependent on the degree of feeding niche overlap. Additionally increasing the degree of feeding specialization of the predators on the two prey species increased the amount of parameter space in which coexistence of the two predators occurred. Bifurcation analyses supported the general pattern of increased stability when the predator interaction was synergistic and decreased stability when it was antagonistic. Thus, synergistic interactions should be more common than antagonistic interactions in ecological systems.     

Antagonistic and bacteriocinogenic activity of Meningococci]

Antagonistic and bacteriocinogenic activity was studied in 169 strains of meningococci of various serological groups and with different localization in the human organism at the time of isolation. Bacteriocinogenic activity was revealed in all the meningococcus strains (by delayed antagonism method), and, in addition, antagonistic activity was found in 100 strains. The inhibitory activity was the greatest in meningococci isolated from the cerebrospinal fluid. The data obtained suggest an important inhibitory activity of meningococci, along with their resistance to the antagonistic activity of the nasopharyngeal microorganisms, in the manifestation of pathogenic properties in them.