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101.
若尔盖高寒湿地干湿土壤条件下微生物群落结构特征 总被引:18,自引:4,他引:14
土壤水分含量的空间异质性是引起湿地生态系统结构和功能空间变异的关键因素。目前有关低纬度高寒湿地土壤水分对微生物群落结构影响的研究较少。于2007年4月(冷季)和8月(暖季)采集若尔盖高寒湿地常年淹水和无淹水两种水分条件的土壤样品,利用磷脂脂肪酸方法分析其微生物群落结构。结果表明,土壤微生物总生物量、细菌生物量、革兰氏阳性细菌及革兰氏阴性细菌生物量均表现为常年淹水土壤高于无淹水土壤,且4月份高于8月份;与土壤通气量关系密切的真菌、放线菌,其生物量表现为无淹水土壤显著高于常年淹水土壤;反映群落组成的真菌:细菌磷脂脂肪酸比值也表现为无淹水土壤显著高于常年淹水土壤。磷脂脂肪酸的主成分分析表明,水分条件不同的两种土壤中微生物群落结构显著不同,季节变化并未引起土壤微生物群落结构的改变。 相似文献
102.
Ashrafali M. Ismail Amrita Saha Ji S. Lee David F. Painter Yinghua Chen Gurdeep Singh Gabriela N. Condezo James Chodosh Carmen San Martín Jaya Rajaiya 《PLoS pathogens》2022,18(6)
As intracellular parasites, viruses exploit cellular proteins at every stage of infection. Adenovirus outbreaks are associated with severe acute respiratory illnesses and conjunctivitis, with no specific antiviral therapy available. An adenoviral vaccine based on human adenovirus species D (HAdV-D) is currently in use for COVID-19. Herein, we investigate host interactions of HAdV-D type 37 (HAdV-D37) protein IIIa (pIIIa), identified by affinity purification and mass spectrometry (AP-MS) screens. We demonstrate that viral pIIIa interacts with ubiquitin-specific protease 9x (USP9x) and Ran-binding protein 2 (RANBP2). USP9x binding did not invoke its signature deubiquitination function but rather deregulated pIIIa-RANBP2 interactions. In USP9x-knockout cells, viral genome replication and viral protein expression increased compared to wild type cells, supporting a host-favored mechanism for USP9x. Conversely, RANBP2-knock down reduced pIIIa transport to the nucleus, viral genome replication, and viral protein expression. Also, RANBP2-siRNA pretreated cells appeared to contain fewer mature viral particles. Transmission electron microscopy of USP9x-siRNA pretreated, virus-infected cells revealed larger than typical paracrystalline viral arrays. RANBP2-siRNA pretreatment led to the accumulation of defective assembly products at an early maturation stage. CRM1 nuclear export blockade by leptomycin B led to the retention of pIIIa within cell nuclei and hindered pIIIa-RANBP2 interactions. In-vitro binding analyses indicated that USP9x and RANBP2 bind to C-terminus of pIIIa amino acids 386–563 and 386–510, respectively. Surface plasmon resonance testing showed direct pIIIa interaction with recombinant USP9x and RANBP2 proteins, without competition. Using an alternative and genetically disparate adenovirus type (HAdV-C5), we show that the demonstrated pIIIa interaction is also important for a severe respiratory pathogen. Together, our results suggest that pIIIa hijacks RANBP2 for nuclear import and subsequent virion assembly. USP9x counteracts this interaction and negatively regulates virion synthesis. This analysis extends the scope of known adenovirus-host interactions and has potential implications in designing new antiviral therapeutics. 相似文献
103.
长白山暗针叶林苔藓植物生物量的研究 总被引:9,自引:0,他引:9
在长白山北坡暗针叶林对地面和树附生苔藓植物的生物量进行了测定.地面生苔藓采取样带调查取样法测定,树附生苔藓应用McCune方法对树干和树枝的附生苔藓生物量都做了细致的测定.结果表明,长白山暗针叶林中的苔藓植物分布很不均匀,随海拔变化差异很大,海拔1100m最低,仅为543kg·hm^-2;海拔1250m最高,达5097kg·hm^-2.苔藓植物生物量的变化对生境有很好的指示作用,特别是塔藓和拟垂枝藓的生物量随海拔的变化与森林系统的群落学特点有一定的相关性:在海拔1100~1700m,塔藓的生物量与臭冷杉的重要值变化趋势相近,随海拔升高而减少;拟垂枝藓的生物量与鱼鳞云杉重要值的变化趋势相似,随海拔升高而增加.此外,生物量随海拔的变化表明了不同苔藓植物对环境条件要求的差异,拟垂枝藓比塔藓水分条件要求更高.因此,生物量的研究在植物生理上也有一定的指示作用. 相似文献
104.
Infection of human dendritic cells by a sindbis virus replicon vector is determined by a single amino acid substitution in the E2 glycoprotein 总被引:5,自引:0,他引:5
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Gardner JP Frolov I Perri S Ji Y MacKichan ML zur Megede J Chen M Belli BA Driver DA Sherrill S Greer CE Otten GR Barnett SW Liu MA Dubensky TW Polo JM 《Journal of virology》2000,74(24):11849-11857
The ability to target antigen-presenting cells with vectors encoding desired antigens holds the promise of potent prophylactic and therapeutic vaccines for infectious diseases and cancer. Toward this goal, we derived variants of the prototype alphavirus, Sindbis virus (SIN), with differential abilities to infect human dendritic cells. Cloning and sequencing of the SIN variant genomes revealed that the genetic determinant for human dendritic cell (DC) tropism mapped to a single amino acid substitution at residue 160 of the envelope glycoprotein E2. Packaging of SIN replicon vectors with the E2 glycoprotein from a DC-tropic variant conferred a similar ability to efficiently infect immature human DC, whereupon those DC were observed to undergo rapid activation and maturation. The SIN replicon particles infected skin-resident mouse DC in vivo, which subsequently migrated to the draining lymph nodes and upregulated cell surface expression of major histocompatibility complex and costimulatory molecules. Furthermore, SIN replicon particles encoding human immunodeficiency virus type 1 p55(Gag) elicited robust Gag-specific T-cell responses in vitro and in vivo, demonstrating that infected DC maintained their ability to process and present replicon-encoded antigen. Interestingly, human and mouse DC were differentially infected by selected SIN variants, suggesting differences in receptor expression between human and murine DC. Taken together, these data illustrate the tremendous potential of using a directed approach in generating alphavirus vaccine vectors that target and activate antigen-presenting cells, resulting in robust antigen-specific immune responses. 相似文献
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108.
用差异显示PCR法筛选与血管外膜细胞表型转化相关的基因 总被引:6,自引:2,他引:6
为筛选血管外膜成纤维细胞(adventitial fibroblast,AF)与肌成纤维细胞(myofibroblast,MF)间表型转化有关的基因,实验建立了大鼠胸主动脉AF和MF两种细胞模型,用差异显示聚合酶链反应(DD-PCR)技术获得表达差异片段,对差异片段进行克隆和测序分析,并用定量PCR和Northern blot对差别显示结果进行验证。用反义核酸转染技术观察骨桥蛋白(osteopontin,OPN)对AF迁移的影响。结果表明,两种表型细胞存在明显的基因表达差异,其中一个在MF下调的差异片段与GenBank中NADH脱氢酶亚单位5(NADH dehydrogenase subunit 5,Nd5)基因高度同源。另一个在MF上调的差异片段与OPN基因同源。上述差异表达结果被定量PCR及Northern blot证实。此外还有4个表达序列标志(expressed sequence-tag,EST)在GenBank中未查到同源序列。反义OPN寡脱氧核甘酸可抑制AF的迁移活动。结果提示,AF转化为MF可能与ND5基因下调、OPN上调及其它未知基因的表达改变有关。应用反义技术适度抑制OPN表达在防治血管重塑中具有重要作用。 相似文献
109.
定向进化提高嗜热拟青霉J18耐热β-1,3-1,4-葡聚糖酶在酸性条件下的催化能力 总被引:1,自引:0,他引:1
应用定向进化技术提高了嗜热拟青霉Paecilomyces thermophila J18耐热β-1,3-1,4-葡聚糖酶(PtLic16A)在酸性条件下的催化能力.结合易错PCR和DNA改组的方法,构建了β-葡聚糖酶的突变体文库;利用刚果红染色法建立了阳性克隆的高通量筛选体系.筛选得到的突变酶PtLic 16AM1的反应最适pH由7.0变化至5.5,且保持了原有的耐热性和比酶活.突变酶的DNA序列中有4个点位发生突变,引发了4处氨基酸替换,分别是T58S、Y110N、G195E和D221G.结构模拟结果显示,发生突变的4个氨基酸位点中,Y110N位置靠近酶活性中心,而T58S、G195E和D221G则离酶活性中心较远,其中T58S、G195E可能对酶最适pH的变化起到了关键作用. 相似文献
110.
Genome-Wide Association Studies in an Isolated Founder Population from the Pacific Island of Kosrae 总被引:1,自引:0,他引:1
Jennifer K. Lowe Julian B. Maller Itsik Pe'er Benjamin M. Neale Jacqueline Salit Eimear E. Kenny Jessica L. Shea Ralph Burkhardt J. Gustav Smith Weizhen Ji Martha Noel Jia Nee Foo Maude L. Blundell Vita Skilling Laura Garcia Marcia L. Sullivan Heather E. Lee Anna Labek Hope Ferdowsian Steven B. Auerbach Richard P. Lifton Christopher Newton-Cheh Jan L. Breslow Markus Stoffel Mark J. Daly David M. Altshuler Jeffrey M. Friedman 《PLoS genetics》2009,5(2)
It has been argued that the limited genetic diversity and reduced allelic heterogeneity observed in isolated founder populations facilitates discovery of loci contributing to both Mendelian and complex disease. A strong founder effect, severe isolation, and substantial inbreeding have dramatically reduced genetic diversity in natives from the island of Kosrae, Federated States of Micronesia, who exhibit a high prevalence of obesity and other metabolic disorders. We hypothesized that genetic drift and possibly natural selection on Kosrae might have increased the frequency of previously rare genetic variants with relatively large effects, making these alleles readily detectable in genome-wide association analysis. However, mapping in large, inbred cohorts introduces analytic challenges, as extensive relatedness between subjects violates the assumptions of independence upon which traditional association test statistics are based. We performed genome-wide association analysis for 15 quantitative traits in 2,906 members of the Kosrae population, using novel approaches to manage the extreme relatedness in the sample. As positive controls, we observe association to known loci for plasma cholesterol, triglycerides, and C-reactive protein and to a compelling candidate loci for thyroid stimulating hormone and fasting plasma glucose. We show that our study is well powered to detect common alleles explaining ≥5% phenotypic variance. However, no such large effects were observed with genome-wide significance, arguing that even in such a severely inbred population, common alleles typically have modest effects. Finally, we show that a majority of common variants discovered in Caucasians have indistinguishable effect sizes on Kosrae, despite the major differences in population genetics and environment. 相似文献