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121.
【目的】利用筛选培养基,从肉牛瘤胃液中分离筛选产乙酰酯酶的细菌菌株,并研究菌株的产酶特征。【方法】利用厌氧培养技术,以木质素为唯一碳源,筛选并驯化所得菌株。根据菌株16S rDNA序列分析、革兰氏染色、伊红美蓝培养基培养、甲基红试验和柠檬酸盐利用试验,鉴定菌株。采用对-硝基苯乙酯测定酶活力。【结果】筛选得到产乙酰酯酶活力较高细菌菌株RB1,初步鉴定为Escherichia coli。菌株RB1的生长曲线表明,0 42 h为菌株的延迟期,42 60 h为菌株的对数期,60 66 h为菌株的稳定期,66 86 h为菌株的衰亡期。菌株所产乙酰酯酶最适温度为40°C,最适pH为8.0,在最适温度与pH条件下,培养基中添加玉米秸秆粉,乙酰酯酶最高酶活力达到0.52 U/mL。【结论】筛选获得产乙酰酯酶的细菌菌株RB1,其乙酰酯酶活力高于已报道的菌株,是一株具有研究和应用潜力的产乙酰酯酶的菌株。 相似文献
122.
The aim of the present study was to understand the effects of abiotic conditions on seasonal feeding activity of diverse herbivores on the same oak tree species in two different forests. We tracked changes in herbivore feeding activities on an oak tree species (Quercus serrata) in two localities: a low elevation small hillock forest patch (Muan, MN) and a middle elevation mountain forest patch (Mt. Jirisan, JR). A total of five sites were selected in each of two forest localities. Data for leaf expansion, leaf chemical qualities, leaf damage ratio, and numbers of lepidopteran caterpillars were collected during spring (May) and summer (July to August), 2012. Leaf expansion rate was higher at the low hillock forest than the mid‐mountain forest from spring to summer. Nitrogen and carbon content decreased seasonally at both localities. Lepidopteran larval diversity was high in the mid‐mountain forest, and two‐way ANOVA showed that species richness of lepidopteran larvae was significantly affected by the interaction between season and locality. Leaf damage by all herbivores was higher in the low hillock forest than the mid‐mountain forest in spring, but was higher in the mid‐mountain forest in summer. Relative proportion of general herbivores increased from spring to summer in the mid‐mountain forest, but not in the low hillock forest. Canonical Correspondence Analysis (CCA) ordination showed that altitude‐ and season‐related variables were significant species and environment interaction factors. Our data indicate that locality and temperature disproportionally affected the feeding activities of diverse herbivores in two different temperate forests. 相似文献
123.
The biochemical mechanisms underlying thidiazuron (TDZ)-induced regeneration in plant cells have not been clearly elucidated.
Exposure of leaf explants of Echinacea purpurea to a medium containing TDZ results in undifferentiated cell proliferation and differentiated growth as mixed shoot organogenesis
and somatic embryogenesis. The current studies were undertaken to determine the potential roles of auxin, indoleamines, and
ion signaling in the dedifferentiation and redifferentiation of plant cells. E. purpurea leaf explants were found to contain auxin and the related indoleamine neurotransmitters, melatonin, and serotonin. The levels
of these endogenous indoleamines were increased by exposure to TDZ associated with the induction of regeneration. The auxin-transport
inhibitor 2,3,5-triiodobenzoic acid and auxin action inhibitor, p-chlorophenoxyisobutyric acid decreased the TDZ-induced regeneration but increased concentrations of endogenous serotonin
and melatonin. As well, inhibitors of calcium and sodium transport significantly reduced TDZ-induced morphogenesis while increasing
endogenous indoleamine content. These data indicate that TDZ-induced regeneration is the manifestation of a metabolic cascade
that includes an initial signaling event, accumulation, and transport of endogenous plant signals such as auxin and melatonin,
a system of secondary messengers, and a concurrent stress response. 相似文献
124.
Hea Jin Ryu Ji-Eun Kim Yeon-Joo Kim Ji-Yang Kim Won IL Kim So-Yeon Choi Min-Ju Kim Tae-Cheon Kang 《Cellular and molecular neurobiology》2013,33(4):575-585
Transient receptor potential canonical channel (TRPC) is a nonselective cation channel permeable to Ca2+, which express in many cell types, including neurons. However the alterations in TRPC receptor expressions in response to status epilepticus (SE) have not been explored. Therefore, the present study was designated to elucidate the roles of TRPC3 in neuronal death and vasogenic edema within the rat piriform cortex (PC) following SE. In non-SE animals, TRPC3 immunoreactivity was abundantly detected in the PC. Following SE, TRPC3 immunoreactivity was increased in neurons. Furthermore, TRPC3 expression was detected in endothelial cells that did not contain it in non-SE animals. Loss of SMI-71 (a blood–brain barrier antigen) immunoreactivity was also observed in TRPC3 positive endothelial cells. In addition, FJB positive neurons and vasogenic edema were noticeably detected in the PC. To directly determine whether TRPC3 activation is correlated to SE-induced vasogenic edema formation and neuronal damages in the PC, the effect of Pyr-3 (a TRPC3 antagonist) on SE-induced insults were investigated. Pyr-3 infusion effectively attenuated vasogenic edema in the PC as compared to the vehicle. Therefore, our findings indicate that TRPC3 activation/overexpression induced by SE may involve BBB disruption and neuronal damages in the rat PC following SE. Therefore, the present study was TRPC3 may play an important role in SE-induced vasogenic edema formation through BBB disruptions in the rat PC. 相似文献
125.
Yung-Tsi Bolon Bindu Joseph Steven B Cannon Michelle A Graham Brian W Diers Andrew D Farmer Gregory D May Gary J Muehlbauer James E Specht Zheng Jin Tu Nathan Weeks Wayne W Xu Randy C Shoemaker Carroll P Vance 《BMC plant biology》2010,10(1):1-24
Background
Cotton fibers (produced by Gossypium species) are the premier natural fibers for textile production. The two tetraploid species, G. barbadense (Gb) and G. hirsutum (Gh), differ significantly in their fiber properties, the former having much longer, finer and stronger fibers that are highly prized. A better understanding of the genetics and underlying biological causes of these differences will aid further improvement of cotton quality through breeding and biotechnology. We evaluated an inter-specific Gh × Gb recombinant inbred line (RIL) population for fiber characteristics in 11 independent experiments under field and glasshouse conditions. Sites were located on 4 continents and 5 countries and some locations were analyzed over multiple years.Results
The RIL population displayed a large variability for all major fiber traits. QTL analyses were performed on a per-site basis by composite interval mapping. Among the 651 putative QTLs (LOD > 2), 167 had a LOD exceeding permutation based thresholds. Coincidence in QTL location across data sets was assessed for the fiber trait categories strength, elongation, length, length uniformity, fineness/maturity, and color. A meta-analysis of more than a thousand putative QTLs was conducted with MetaQTL software to integrate QTL data from the RIL and 3 backcross populations (from the same parents) and to compare them with the literature. Although the global level of congruence across experiments and populations was generally moderate, the QTL clustering was possible for 30 trait x chromosome combinations (5 traits in 19 different chromosomes) where an effective co-localization of unidirectional (similar sign of additivity) QTLs from at least 5 different data sets was observed. Most consistent meta-clusters were identified for fiber color on chromosomes c6, c8 and c25, fineness on c15, and fiber length on c3.Conclusions
Meta-analysis provided a reliable means of integrating phenotypic and genetic mapping data across multiple populations and environments for complex fiber traits. The consistent chromosomal regions contributing to fiber quality traits constitute good candidates for the further dissection of the genetic and genomic factors underlying important fiber characteristics, and for marker-assisted selection. 相似文献126.
Williams DC Wildung MR Jin AQ Dalal D Oliver JS Coates RM Croteau R 《Archives of biochemistry and biophysics》2000,379(1):137-146
The diterpene cyclase taxadiene synthase from yew (Taxus) species transforms geranylgeranyl diphosphate to taxa-4(5),11(12)-diene as the first committed step in the biosynthesis of the anti-cancer drug Taxol. Taxadiene synthase is translated as a preprotein bearing an N-terminal targeting sequence for localization to and processing in the plastids. Overexpression of the full-length preprotein in Escherichia coli and purification are compromised by host codon usage, inclusion body formation, and association with host chaperones, and the preprotein is catalytically impaired. Since the transit peptide-mature enzyme cleavage site could not be determined directly, a series of N-terminally truncated enzymes was created by expression of the corresponding cDNAs from a suitable vector, and each was purified and kinetically evaluated. Deletion of up to 79 residues yielded functional protein; however, deletion of 93 or more amino acids resulted in complete elimination of activity, implying a structural or catalytic role for the amino terminus. The pseudomature form of taxadiene synthase having 60 amino acids deleted from the preprotein was found to be superior with respect to level of expression, ease of purification, solubility, stability, and catalytic activity with kinetics comparable to the native enzyme. In addition to the major product, taxa-4(5),11(12)-diene (94%), this enzyme produces a small amount of the isomeric taxa-4(20), 11(12)-diene ( approximately 5%), and a product tentatively identified as verticillene ( approximately 1%). Isotopically sensitive branching experiments utilizing (4R)-[4-(2)H(1)]geranylgeranyl diphosphate confirmed that the two taxadiene isomers, and a third (taxa-3(4),11(12)-diene), are derived from the same intermediate taxenyl C4-carbocation. These results, along with the failure of the enzyme to utilize 2, 7-cyclogeranylgeranyl diphosphate as an alternate substrate, indicate that the reaction proceeds by initial ionization of the diphosphate ester and macrocyclization to the verticillyl intermediate, followed by a secondary cyclization to the taxenyl cation and deprotonation (i.e., formation of the A-ring prior to B/C-ring closure). Two potential mechanism-based inhibitors were tested with recombinant taxadiene synthase but neither provided time-dependent inactivation nor afforded more than modest competitive inhibition. 相似文献
127.
人乳头瘤病毒分型检测在宫颈病变中的研究进展 总被引:1,自引:0,他引:1
宫颈癌是感染性疾病,HPV感染是宫颈癌发生的主要因素。宫颈上皮内留样病变(CIN)是宫颈浸润癌演变发展过程中的癌前病变阶段,研究发现大多数CIN伴有人乳头瘤病毒(HPV)感染,不同HPV亚型的致病力不同,因此不同亚型的HPV感染可以导致不同的宫颈病变。对CIN早诊、早治是降低宫颈癌发生率和死亡率的关键,临床上应用HPV亚型检测对宫颈病变的初筛及治疗追踪具有重要意义。 相似文献
128.
Reagentless amperometric immunosensor for human chorionic gonadotrophin based on direct electrochemistry of horseradish peroxidase 总被引:4,自引:0,他引:4
A novel amperometric immunosensor for determination of human serum chorionic gonadotrophin (HCG) was constructed by immobilization of HCG with titania sol-gel on a glassy carbon electrode and the direct electrochemistry of horseradish peroxidase (HRP) labeled to HCG antibody (HRP-anti-HCG). The morphologies of the HCG membrane were characterized to be chemically clean, porous and homogeneous. HRP-anti-HCG was functionally conjugated with the immobilized HCG after incubation in phosphate buffer (PBS) containing HRP-anti-HCG. A direct electron transfer of HRP with a rate constant of 1.35+/-0.40 s(-1) was observed at the HRP-anti-HCG-HCG/titania sol-gel membrane modified electrode in 0.1 M PBS pH 7.0. With a competitive mechanism the differential pulse voltammetric peak current of the immobilized HRP decreased linearly with an increasing HCG concentration from 2.5 to 12.5 mIU/ml in the incubation solution. The HCG immunosensor showed a detection limit of 1.4 mIU/ml, a good accuracy and acceptable precision and reproducibility with an intra-assay CV of 4.7% at 5.0 mIU/ml and an inter-assay precision of 8.1% obtained at 10 mIU/ml. The biosensor displayed a good stability in a storage period of 30 days. 相似文献
129.
人Leptin基因在大肠杆菌中的高效表达、纯化及其生物学活性研究 总被引:5,自引:0,他引:5
将人Leptin表达质粒pBV220-OB转化E.coliJM109,经热诱导获得了目的蛋白的表达。经SDS-PAGE鉴定分析,表达产物以包涵体形式存在,目的蛋白表达量占菌体总蛋白的40%以上。通过包涵体分离,Sephacryl S200HR凝胶和DEAE52离子交换层析及Hypersil C18柱反相色谱纯化,获得纯度在95%以上,内毒素含量小于10EU/mg的高纯度的重组人Leptin。Western-blot鉴定表明,纯化表达产物能和抗Leptin抗体特异性结合;蛋白质N端15个氨基酸序列分析结果和预期的序列一致。纯化产物经复性处理,其分子中Cys96和Cys146形成二硫键。体内活性检测显示,纯化和复性的rh-Leptin明显抑制BALB/c小鼠的进食和体重增长,提示其具有明显的生物学活性。 相似文献
130.
Loren P. Albert Natalia Restrepo‐Coupe Marielle N. Smith Jin Wu Cecilia Chavana‐Bryant Neill Prohaska Tyeen C. Taylor Giordane A. Martins Philippe Ciais Jiafu Mao M. Altaf Arain Wei Li Xiaoying Shi Daniel M. Ricciuto Travis E. Huxman Sean M. McMahon Scott R. Saleska 《Global Change Biology》2019,25(11):3591-3608
Plant phenology—the timing of cyclic or recurrent biological events in plants—offers insight into the ecology, evolution, and seasonality of plant‐mediated ecosystem processes. Traditionally studied phenologies are readily apparent, such as flowering events, germination timing, and season‐initiating budbreak. However, a broad range of phenologies that are fundamental to the ecology and evolution of plants, and to global biogeochemical cycles and climate change predictions, have been neglected because they are “cryptic”—that is, hidden from view (e.g., root production) or difficult to distinguish and interpret based on common measurements at typical scales of examination (e.g., leaf turnover in evergreen forests). We illustrate how capturing cryptic phenology can advance scientific understanding with two case studies: wood phenology in a deciduous forest of the northeastern USA and leaf phenology in tropical evergreen forests of Amazonia. Drawing on these case studies and other literature, we argue that conceptualizing and characterizing cryptic plant phenology is needed for understanding and accurate prediction at many scales from organisms to ecosystems. We recommend avenues of empirical and modeling research to accelerate discovery of cryptic phenological patterns, to understand their causes and consequences, and to represent these processes in terrestrial biosphere models. 相似文献