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  • 文章类型: Journal Article
    Group-wise diversity of sediment methylotrophs of Chilika lake (Lat. 19°28\'-19°54\'N; Long. 85°06\'-85°35\'E) Odisha, India at various identified sites was studied. Both the culturable and unculturable (metagenome) methylotrophs were investigated in the lake sediments employing both mxaF and 16S rRNA genes as markers. ARDRA profiling, 16S rRNA gene sequencing, PAGE profiling of HaeIII, EcoRI restricted mxaF gene and the mxaF gene sequences using culture-dependent approach revealed the relatedness of α-proteobacteria and Methylobacterium, Hyphomicrobium and Ancyclobacter sp. The total viable counts of the culturable aerobic methylotrophs were relatively higher in sediments near the sea mouth (S3; Panaspada), also demonstrated relatively high salinity (0.1 M NaCl) tolerance. Metagenomic DNA from the sediments, amplified using GC clamp mxaF primers and resolved through DGGE, revealed the diversity within the unculturable methylotrophic bacterium Methylobacterium organophilum, Ancyclobacter aquaticus, Burkholderiales and Hyphomicrobium sp. Culture-independent analyses revealed that up to 90 % of the methylotrophs were unculturable. The study enhances the general understandings of the metagenomic methylotrophs from such a special ecological niche.
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  • 文章类型: Comparative Study
    复杂蛋白质组样品的深入分析需要在shot弹枪蛋白质组学实验中的LC-MS/MS之前将样品分级分离为子样品。我们已经建立了shot弹枪蛋白质组学的3D工作流程,该流程依赖于通过1DPAGE进行蛋白质分离,凝胶分馏,胰蛋白酶消化,和通过凝胶内IEF分离肽,在RP-HPLC-MS/MS之前我们的结果表明,应用IEF肽可以显着增加从PAGE亚分级分离鉴定的蛋白质数量。该方法提供了更深的蛋白质组覆盖,并通过仍然依赖于根据第一维分子量的蛋白质分离,在实验上接近高度复杂的混合物时提供了很大程度的灵活性。
    The in-depth analysis of complex proteome samples requires fractionation of the sample into subsamples prior to LC-MS/MS in shotgun proteomics experiments. We have established a 3D workflow for shotgun proteomics that relies on protein separation by 1D PAGE, gel fractionation, trypsin digestion, and peptide separation by in-gel IEF, prior to RP-HPLC-MS/MS. Our results show that applying peptide IEF can significantly increase the number of proteins identified from PAGE subfractionation. This method delivers deeper proteome coverage and provides a large degree of flexibility in experimentally approaching highly complex mixtures by still relying on protein separation according to molecular weight in the first dimension.
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  • 文章类型: Journal Article
    许多影响血浆高密度脂蛋白胆固醇的常见遗传变异,低密度脂蛋白胆固醇(LDL-C),和甘油三酯分布已通过全基因组关联研究(GWAS)确定。然而,这些关联是否与年龄相关,在很大程度上被忽视了。我们在22,000多名欧裔美国人中进行了一项关联研究和荟萃分析,其中包括49种先前鉴定的GWAS变体和三种脂质性状,按年龄分层(男性:<50岁或≥50岁;女性:绝经前或绝经后)。对于每个变体,在两个年龄层次之间进行异质性检验,并使用显著的Phet值作为年龄特异性遗传效应的证据.我们确定了女性中的七个关联和男性中的八个关联,这些关联显示出年龄的暗示性异质性(Phet<0.05)。男性rs174547(FADS1)和LDL-C之间的关联显示出年龄组之间异质性的最多证据(Phet=1.74E-03,I(2)=89.8),与老年男性(P=1.39E-06)显着相关,但与年轻男性(P=0.99)无关。然而,没有一个暗示性修饰效应在多重测试的调整中幸存下来,突出了尽管样本量很大,但识别适度SNP-性状关联的修饰符的挑战。
    Numerous common genetic variants that influence plasma high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglyceride distributions have been identified via genome-wide association studies (GWAS). However, whether or not these associations are age-dependent has largely been overlooked. We conducted an association study and meta-analysis in more than 22,000 European Americans between 49 previously identified GWAS variants and the three lipid traits, stratified by age (males: <50 or ≥50 years of age; females: pre- or postmenopausal). For each variant, a test of heterogeneity was performed between the two age strata and significant Phet values were used as evidence of age-specific genetic effects. We identified seven associations in females and eight in males that displayed suggestive heterogeneity by age (Phet < 0.05). The association between rs174547 (FADS1) and LDL-C in males displayed the most evidence for heterogeneity between age groups (Phet = 1.74E-03, I(2) = 89.8), with a significant association in older males (P = 1.39E-06) but not younger males (P = 0.99). However, none of the suggestive modifying effects survived adjustment for multiple testing, highlighting the challenges of identifying modifiers of modest SNP-trait associations despite large sample sizes.
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