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1.
Optimizing antibody immobilization strategies for the construction of protein microarrays 总被引:11,自引:0,他引:11
Peluso P Wilson DS Do D Tran H Venkatasubbaiah M Quincy D Heidecker B Poindexter K Tolani N Phelan M Witte K Jung LS Wagner P Nock S 《Analytical biochemistry》2003,312(2):113-124
Antibody microarrays have the potential to revolutionize protein expression profiling. The intensity of specific signal produced on a feature of such an array is related to the amount of analyte that is captured from the biological mixture by the immobilized antibody (the "capture agent"). This in turn is a function of the surface density and fractional activity of the capture agents. Here we investigate how these two factors are affected by the orientation of the capture agents on the surface. We compare randomly versus specifically oriented capture agents based on both full-sized antibodies and Fab' fragments. Each comparison was performed using three different antibodies and two types of streptavidin-coated monolayer surfaces. The specific orientation of capture agents consistently increases the analyte-binding capacity of the surfaces, with up to 10-fold improvements over surfaces with randomly oriented capture agents. Surface plasmon resonance revealed a dense monolayer of Fab' fragments that are on average 90% active when specifically oriented. Randomly attached Fab's could not be packed at such a high density and generally also had a lower specific activity. These results emphasize the importance of attaching proteins to surfaces such that their binding sites are oriented toward the solution phase. 相似文献
2.
Dorian?FullerEmail author Ravi?Korisettar P.?C.?Venkatasubbaiah Martin?K.?Jones 《Vegetation History and Archaeobotany》2004,13(2):115-129
Analysis of flotation samples from twelve sites in Karnataka and Andhra Pradesh (south India) provides clear evidence for the predominant subsistence plants of the Neolithic period (2,800–1,200 cal b.c.). This evidence indicates that the likely staples were two pulses (Vigna radiata and Macrotyloma uniflorum) and two millet-grasses (Brachiaria ramosa and Setaria verticillata) which were indigenous to the Indian peninsula. At some sites there is evidence for limited cultivation of wheats (Triticum diococcum, Triticum durum/aestivum) and barley (Hordeum vulgare), and a few crops that originated in Africa, including hyacinth bean (Lablab purpureus), pearl millet (Pennisetum glaucum) and finger millet (Eleusine coracana). In addition there is evidence for cotton (Gossypium sp.), and linseed (Linum sp.), as well as gathered fruits of Ziziphus and two Cucurbitaceae. This evidence suggests that the earliest agriculture in south India, dating to the third millennium b.c., was based on plants domesticated in the region, and that subsequently from the late 3rd millennium b.c. through the 2nd millennium additional crops from other regions were adopted into the subsistence system.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00334-004-0036-9 相似文献
3.
Clio Der Sarkissian Morten E. Allentoft María C. ávila-Arcos Ross Barnett Paula F. Campos Enrico Cappellini Luca Ermini Ruth Fernández Rute da Fonseca Aurélien Ginolhac Anders J. Hansen Hákon Jónsson Thorfinn Korneliussen Ashot Margaryan Michael D. Martin J. Víctor Moreno-Mayar Maanasa Raghavan Morten Rasmussen Marcela Sandoval Velasco Hannes Schroeder Mikkel Schubert Andaine Seguin-Orlando Nathan Wales M. Thomas P. Gilbert Eske Willerslev Ludovic Orlando 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2015,370(1660)
The past decade has witnessed a revolution in ancient DNA (aDNA) research. Although the field''s focus was previously limited to mitochondrial DNA and a few nuclear markers, whole genome sequences from the deep past can now be retrieved. This breakthrough is tightly connected to the massive sequence throughput of next generation sequencing platforms and the ability to target short and degraded DNA molecules. Many ancient specimens previously unsuitable for DNA analyses because of extensive degradation can now successfully be used as source materials. Additionally, the analytical power obtained by increasing the number of sequence reads to billions effectively means that contamination issues that have haunted aDNA research for decades, particularly in human studies, can now be efficiently and confidently quantified. At present, whole genomes have been sequenced from ancient anatomically modern humans, archaic hominins, ancient pathogens and megafaunal species. Those have revealed important functional and phenotypic information, as well as unexpected adaptation, migration and admixture patterns. As such, the field of aDNA has entered the new era of genomics and has provided valuable information when testing specific hypotheses related to the past. 相似文献
4.
Rashmi Venkatasubbaiah Prabhushankar Hema Bommanamane Sanjay Konasur Rajesh 《Plant Cell, Tissue and Organ Culture》2021,146(3):515-529
Plant Cell, Tissue and Organ Culture (PCTOC) - This study proposes the tenacious vitality for biomass production and eco-friendly, cost-effective, renewable silver nanoparticles (AgNPs)... 相似文献
5.
Anna Juras Miroslawa Dabert Alena Kushniarevich Helena Malmstr?m Maanasa Raghavan Jakub Z. Kosicki Ene Metspalu Eske Willerslev Janusz Piontek 《PloS one》2014,9(10)
While numerous ancient human DNA datasets from across Europe have been published till date, modern-day Poland in particular, remains uninvestigated. Besides application in the reconstruction of continent-wide human history, data from this region would also contribute towards our understanding of the history of the Slavs, whose origin is hypothesized to be in East or Central Europe. Here, we present the first population-scale ancient human DNA study from the region of modern-day Poland by establishing mitochondrial DNA profiles for 23 samples dated to 200 BC – 500 AD (Roman Iron Age) and for 20 samples dated to 1000–1400 AD (Medieval Age). Our results show that mitochondrial DNA sequences from both periods belong to haplogroups that are characteristic of contemporary West Eurasia. Haplotype sharing analysis indicates that majority of the ancient haplotypes are widespread in some modern Europeans, including Poles. Notably, the Roman Iron Age samples share more rare haplotypes with Central and Northeast Europeans, whereas the Medieval Age samples share more rare haplotypes with East-Central and South-East Europeans, primarily Slavic populations. Our data demonstrates genetic continuity of certain matrilineages (H5a1 and N1a1a2) in the area of present-day Poland from at least the Roman Iron Age until present. As such, the maternal gene pool of present-day Poles, Czechs and Slovaks, categorized as Western Slavs, is likely to have descended from inhabitants of East-Central Europe during the Roman Iron Age. 相似文献
6.
Julia T. Vilstrup Andaine Seguin-Orlando Mathias Stiller Aurelien Ginolhac Maanasa Raghavan Sandra C. A. Nielsen Jacobo Weinstock Duane Froese Sergei K. Vasiliev Nikolai D. Ovodov Joel Clary Kristofer M. Helgen Robert C. Fleischer Alan Cooper Beth Shapiro Ludovic Orlando 《PloS one》2013,8(2)
The genus Equus is richly represented in the fossil record, yet our understanding of taxonomic relationships within this genus remains limited. To estimate the phylogenetic relationships among modern horses, zebras, asses and donkeys, we generated the first data set including complete mitochondrial sequences from all seven extant lineages within the genus Equus. Bayesian and Maximum Likelihood phylogenetic inference confirms that zebras are monophyletic within the genus, and the Plains and Grevy’s zebras form a well-supported monophyletic group. Using ancient DNA techniques, we further characterize the complete mitochondrial genomes of three extinct equid lineages (the New World stilt-legged horses, NWSLH; the subgenus Sussemionus; and the Quagga, Equus quagga quagga). Comparisons with extant taxa confirm the NWSLH as being part of the caballines, and the Quagga and Plains zebras as being conspecific. However, the evolutionary relationships among the non-caballine lineages, including the now-extinct subgenus Sussemionus, remain unresolved, most likely due to extremely rapid radiation within this group. The closest living outgroups (rhinos and tapirs) were found to be too phylogenetically distant to calibrate reliable molecular clocks. Additional mitochondrial genome sequence data, including radiocarbon dated ancient equids, will be required before revisiting the exact timing of the lineage radiation leading up to modern equids, which for now were found to have possibly shared a common ancestor as far as up to 4 Million years ago (Mya). 相似文献
7.
Venkatesha VA Venkataraman S Sarsour EH Kalen AL Buettner GR Robertson LW Lehmler HJ Goswami PC 《Free radical biology & medicine》2008,45(8):1094-1102
Polychlorinated biphenyls (PCBs) are environmental chemical contaminants believed to adversely affect cellular processes. We investigated the hypothesis that PCB-induced changes in the levels of cellular reactive oxygen species (ROS) induce DNA damage resulting in cytotoxicity. Exponentially growing cultures of human nonmalignant breast epithelial cells (MCF10A) were incubated with PCBs for 3 days and assayed for cell number, ROS levels, DNA damage, and cytotoxicity. Exposure to 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153) or 2-(4-chlorophenyl)benzo-1,4-quinone (4-Cl-BQ), a metabolite of 4-chlorobiphenyl (PCB3), significantly decreased cell number and MTS reduction and increased the percentage of cells with sub-G1 DNA content. Results from electron paramagnetic resonance (EPR) spectroscopy showed a 4-fold increase in the steady-state levels of ROS, which was suppressed in cells pretreated with catalase. EPR measurements in cells treated with 4-Cl-BQ detected the presence of a semiquinone radical, suggesting that the increased levels of ROS could be due to the redox cycling of 4-Cl-BQ. A dose-dependent increase in micronuclei frequency was observed in PCB-treated cells, consistent with an increase in histone 2AX phosphorylation. Treatment of cells with catalase blunted the PCB-induced increase in micronuclei frequency and H2AX phosphorylation that was consistent with an increase in cell survival. Our results demonstrate a PCB-induced increase in cellular levels of ROS causing DNA damage, resulting in cell killing. 相似文献
8.
Chandrasekhar V Deria P Krishnan V Athimoolam A Singh S Madhavaiah C Srivatsan SG Verma S 《Bioorganic & medicinal chemistry letters》2004,14(6):1559-1562
Pendant pyrazolylcyclophosphazene containing hybrid cross-linked polymer (CPPL) has been utilized for binding Zn(II). The metalated polymer (CPPL-Zn) has been found to be very effective catalyst for the hydrolysis of a RNA model phosphodiester substrate [2-(hydroxypropyl)-p-nitrophenyl phosphate, hNPP]. In addition, CPPL-Zn also cleaved supercoiled plasmid DNA pBR322 thus providing a novel structural motif of inorganic-organic hybrid polymers as synthetic nucleases. 相似文献
9.
Stagonospora apocyni causes a leaf spot disease on hemp dogbane (Apocynum cannabinum L.). The fungus produced phytotoxins citrinin, mellein, tyrosol and α–acetylorcinol in liquid culture. All toxins caused necrosis when placed on leaves of hemp dogbane, and eight other weed species. All four toxins were non–specific phytotoxins. Citrinin showed antimicrobial properties against some bacteria and fungi. 相似文献
10.
Impaired macrophage function underscores susceptibility to Salmonella in mice lacking Irgm1 (LRG-47)
Henry SC Daniell X Indaram M Whitesides JF Sempowski GD Howell D Oliver T Taylor GA 《Journal of immunology (Baltimore, Md. : 1950)》2007,179(10):6963-6972
IRG proteins, or immunity-related GTPases (also known as p47 GTPases), are a group of IFN-regulated proteins that are highly expressed in response to infection. The proteins localize to intracellular membranes including vacuoles that contain pathogens in infected macrophages and other host cells. Current data indicate that the IRG protein Irgm1 (LRG-47) is critical for resistance to intracellular bacteria. This function is thought to be a consequence of regulating the survival of vacuolar bacteria in host cells. In the current work, the role of Irgm1 in controlling resistance to Salmonella typhimurium was explored to further define the mechanism through which the protein regulates host resistance. Irgm1-deficient mice displayed increased susceptibility to this bacterium that was reflected in increased bacterial loads in spleen and liver and decreased maturation of S. typhimurium granulomas. The mice also displayed an inability to concentrate macrophages at sites of bacterial deposition. In vitro, the ability of Irgm1-deficient macrophages to suppress intracellular growth of S. typhimurium was impaired. Furthermore, adhesion and motility of Irgm1-deficient macrophages after activation with IFN-gamma was markedly decreased. Altered adhesion/motility of those cells was accompanied by changes in cell morphology, density of adhesion-associated proteins, and actin staining. Together, these data suggest that in addition to regulating the maturation of pathogen-containing vacuoles, Irgm1 plays a key role in regulating the adhesion and motility of activated macrophages. 相似文献