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281.
Acidic region streaking (ARS) is one of the lacunae in two-dimensional gel electrophoresis (2DE) of bacterial proteome. This streaking is primarily caused by nucleic acid (NuA) contamination and poses major problem in the downstream processes like image analysis and protein identification. Although cleanup and nuclease digestion are practiced as remedial options, these strategies may incur loss in protein recovery and perform incomplete removal of NuA. As a result, ARS has remained a common observation across publications, including the recent ones. In this work, we demonstrate how ultrasound wave can be used to shear NuA in plain ice-cooled water, facilitating the elimination of ARS in the 2DE gels without the need for any additional sample cleanup tasks. In combination with a suitable buffer recipe, IEF program and frequent paper-wick changing approach, we are able to reproducibly demonstrate the production of clean 2DE gels with improved protein recovery and negligible or no ARS. We illustrate our procedure using whole cell protein extracts from two diverse organisms, Escherichia coli and Mycobacterium smegmatis. Our designed protocols are straightforward and expected to provide good 2DE gels without ARS, with comparable times and significantly lower cost. 相似文献
282.
Rupak K. Sarma Animesh Gogoi Budheswar Dehury Rajal Debnath Tarun C. Bora Ratul Saikia 《PloS one》2014,9(10)
Study on microbial diversity in the unexplored rhizosphere is important to understand their community structure, biology and ecological interaction with the host plant. This research assessed the genetic and functional diversity of fluorescent pseudomonads [FP] in the green gram rhizophere. One hundred and twenty types of morphologically distinct fluorescent pseudomonads were isolated during vegetative as well as reproductive growth phase of green gram. Rep PCR, ARDRA and RISA revealed two distinct clusters in each case at 75, 61 and 70% similarity coefficient index respectively. 16S rRNA partial sequencing analysis of 85 distantly related fluorescent pseudomonads depicted Pseudomonas aeruginosa as the dominant group. Out of 120 isolates, 23 (19%) showed antagonistic activity towards phytopathogenic fungi. These bacterial isolates showed varied production of salicylic acid, HCN and chitinase, 2, 4-diacetylphloroglucinol (DAPG), phenazine-1-carboxylic acid (PCA) and pyoluteorin (PLT). Production efficiency of inherent level of plant growth promoting (PGP) traits among the 120 isolates demonstrated that 10 (8%) solubilised inorganic phosphates, 25 (20%) produced indoles and 5 (4%) retained ACC deaminase activity. Pseudomonas aeruginosa GGRJ21 showed the highest production of all antagonistic and plant growth promoting (PGP) traits. In a greenhouse experiment, GGRJ21 suppressed root rot disease of green gram by 28–93% (p = 0.05). Consistent up regulation of three important stress responsive genes, i.e., acdS, KatA and gbsA and elevated production efficiency of different PGP traits could promote GGRJ21 as a potent plant growth regulator. 相似文献
283.
284.
Christopher Arico-Muendel Paolo A. Centrella Brooke D. Contonio Barry A. Morgan Gary O’Donovan Christopher L. Paradise Steven R. Skinner Barbara Sluboski Jennifer L. Svendsen Kerry F. White Anjan Debnath Jiri Gut Nathan Wilson James H. McKerrow Joseph L. DeRisi Philip J. Rosenthal Peter K. Chiang 《Bioorganic & medicinal chemistry letters》2009,19(17):5128-5131
Fumagillin, an irreversible inhibitor of MetAP2, has been shown to potently inhibit growth of malaria parasites in vitro. Here, we demonstrate activity of fumagillin analogs with an improved pharmacokinetic profile against malaria parasites, trypanosomes, and amoebas. A subset of the compounds showed efficacy in a murine malaria model. The observed SAR forms a basis for further optimization of fumagillin based inhibitors against parasitic targets by inhibition of MetAP2. 相似文献
285.
Tim Downing David J. Lynn Sarah Connell Andrew T. Lloyd AK Fazlul Haque Bhuiyan Pradeepa Silva Arifa N. Naqvi Rahamame Sanfo Racine-Samba Sow Baitsi Podisi Cliona O’Farrelly Olivier Hanotte Daniel G. Bradley 《Immunogenetics》2009,61(4):303-314
There have been significant evolutionary pressures on the chicken during both its speciation and its subsequent domestication
by man. Infectious diseases are expected to have exerted strong selective pressures during these processes. Consequently,
it is likely that genes associated with disease susceptibility or resistance have been subject to some form of selection.
Two genes involved in the immune response (interferon-γ and interleukin 1-β) were selected for sequencing in diverse chicken
populations from Pakistan, Sri Lanka, Bangladesh, Kenya, Senegal, Burkina Faso and Botswana, as well as six outgroup samples
(grey, green, red and Ceylon jungle fowl and grey francolin and bamboo partridge). Haplotype frequencies, tests of neutrality,
summary statistics, coalescent simulations and phylogenetic analysis by maximum likelihood were used to determine the population
genetic characteristics of the genes. Networks indicate that these chicken genes are most closely related to the red jungle
fowl. Interferon-γ had lower diversity and considerable coding sequence conservation, which is consistent with its function
as a key inflammatory cytokine of the immune response. In contrast, the pleiotropic cytokine interleukin 1-β had higher diversity
and showed signals of balancing selection moderated by recombination, yielding high numbers of diverse alleles, possibly reflecting
broader functionality and potential roles in more diseases in different environments.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
An erratum to this article can be found at 相似文献
286.
Samir C. Debnath 《Nordic Journal of Botany》2009,27(2):141-148
Understanding of the genetic relationship within wild lowbush blueberry ( Vaccinium angustifolium Ait.) germplasm is important to establish a broad genetic base for safeguarding and for future use of the existing genetic resources. The objective of this study was to assess the genetic variability within 43 wild lowbush blueberry clones, collected from 10 communities of four Canadian provinces, and the cultivar 'Fundy' by using inter simple sequence repeat (ISSR) markers, with the hope to establish a reference set of lowbush blueberry germplasm for blueberry conservation, breeding and research. Thirteen primers generated 242 polymorphic ISSR-PCR bands. A substantial degree of genetic similarity was found among the wild collections. Cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) separated the 41 genotypes into two main clusters, and identified the three remaining clones as outliers. Furthermore, within one main cluster, the genotypes tended to form sub-clusters that were in agreement with the principal coordinate (PCO) analysis. Geographical distribution contributed to 27% of total variation as revealed by analysis of molecular variance (AMOVA). The ISSR-PCR method was simple, fast and relatively inexpensive to produce useful DNA fragments and detected a sufficient degree of polymorphism to differentiate among lowbush blueberry clones, making this technology valuable for germplasm management and the more efficient choice of parents in current blueberry breeding program. 相似文献
287.
F. Marc Michel Hazel-Ann Hosein Douglas B. Hausner Sudeep Debnath John B. Parise Daniel R. Strongin 《Biochimica et Biophysica Acta (BBA)/General Subjects》2010
Background
In nature or in the laboratory, the roughly spherical interior of the ferritin protein is well suited for the formation and storage of a variety of nanosized metal oxy-hydroxide compounds which hold promise for a range of applications. However, the linkages between ferritin reactivity and the structure and physicochemical properties of the nanoparticle core, either native or reconstituted, remain only partly understood.Scope of review
Here we review studies, including those from our laboratory, which have investigated the structure of ferritin-derived ferrihydrite and reactivity of ferritin, both native and reconstituted. Selected proposed structure models for ferrihydrite are discussed along with the structural and genetic relationships that exist among several different forms of ferrihydrite. With regard to reactivity, the review will emphasize studies that have investigated the (photo)reactivity of ferritin and ferritin-derived materials with environmentally relevant gaseous and aqueous species.Major conclusions
The inorganic core formed from apoferritin reconstituted with varied amounts of Fe has the same structural topology as the inorganically derived ferrihydrite that is an important component of many environmental and soil systems. Reactivity of ferritin toward aqueous species resulting from the photoexcitation of the inorganic core of the protein shows promise for driving redox reactions relevant to environmental chemistry.General significance
Ferritin-derived ferrihydrite is effectively maintained in a relatively unaggregated state, which improves reactivity and opens the possibility of future applications in environmental remediation. Advances in our understanding of the structure, composition, and disorder in synthetic, inorganically derived ferrihydrite are shedding new light on the reactivity and stability of ferrihydrite derived artificially from ferritin. 相似文献288.
Lanying Du Guangyu Zhao Zhihua Kou Cuiqing Ma Shihui Sun Vincent K. M. Poon Lu Lu Lili Wang Asim K. Debnath Bo-Jian Zheng Yusen Zhou Shibo Jiang 《Journal of virology》2013,87(17):9939-9942
A novel human Middle East respiratory syndrome coronavirus (MERS-CoV) caused outbreaks of severe acute respiratory syndrome (SARS)-like illness with a high mortality rate, raising concerns of its pandemic potential. Dipeptidyl peptidase-4 (DPP4) was recently identified as its receptor. Here we showed that residues 377 to 662 in the S protein of MERS-CoV specifically bound to DPP4-expressing cells and soluble DPP4 protein and induced significant neutralizing antibody responses, suggesting that this region contains the receptor-binding domain (RBD), which has a potential to be developed as a MERS-CoV vaccine. 相似文献
289.
Molecular cloning and characterization of an alpha1,3 fucosyltransferase, CEFT-1, from Caenorhabditis elegans 总被引:1,自引:1,他引:1
We report on the identification, molecular cloning, and characterization of
an alpha1,3 fucosyltransferase (alpha1,3FT) expressed by the nematode,
Caenorhabditis elegans . Although C. elegans glycoconjugates do not express
the Lewis x antigen Galbeta1-- >4[Fucalpha1-->3]GlcNAcbeta-->R,
detergent extracts of adult C.elegans contain an alpha1,3FT that can
fucosylate both nonsialylated and sialylated acceptor glycans to generate
the Lexand sialyl Lexantigens, as well as the lacdiNAc-containing acceptor
GalNAcbeta1-->4GlcNAcbeta1-- >R to generate GalNAcbeta1-->4
[Fucalpha1-->3]GlcNAcbeta1-->R. A search of the C.elegans genome
database revealed the existence of a gene with 20-23% overall identity to
all five cloned human alpha1,3FTs. The putative cDNA for the C.elegans
alpha1,3FT (CEFT-1) was amplified by PCR from a cDNA lambdaZAP library,
cloned, and sequenced. COS7 cells transiently transfected with cDNA
encoding CEFT-1 express the Lex, but not sLexantigen. The CEFT-1 in the
transfected cell extracts can synthesize Lex, but not sialyl Lex, using
exogenous acceptors. A second fucosyltransferase activity was detected in
extracts of C. elegans that transfers Fuc in alpha1,2 linkage to Gal
specifically on type-1 chains. The discovery of alpha-fucosyltransferases
in C. elegans opens the possibility of using this well-characterized
nematode as a model system for studying the role of fucosylated glycans in
the development and survival of C.elegans and possibly other helminths.
相似文献
290.
ABSTRACT: BACKGROUND: p21-activated kinase (PAK) has been implicated in the inflammatory activation of endothelial cells by disturbed fluid shear stress, which is the initiating stimulus in atherosclerosis. The study addresses whether PAK1 contributes to inflammatory marker expression in endothelial cells at atherosclerosis-susceptible regions of arteries in vivo. METHOD: Aortas from WT and PAK1-/- C57BL/6J mice on a normal chow diet were fixed, dissected and processed for immunohistochemistry using a panel of inflammatory markers. We visualized and quantified staining in the endothelium at the greater and lesser curvatures of the arch of aorta, as atherosclerosis-resistant and susceptible regions, respectively. RESULTS: Fibronectin, VCAM-1 and the activated RelA NF-kappaB subunit were localized to the lesser curvature and decreased in PAK1-/- mice. The activated RelB NF-kappaB subunit was also localized to the lesser curvature but was increased in PAK1-/- mice. Low levels of staining for ICAM-1 and the monocyte/macrophage marker Mac2 indicated that overall inflammation in this tissue was minimal. CONCLUSION: These data show that PAK1 has a significant pro-inflammatory function at atherosclerosisprone sites in vivo. These effects are seen in young mice with very low levels of inflammation, suggesting that inflammatory activation of the endothelium is primarily biomechanical. Activation involves NF-kappaB, expression of leukocyte recruitment receptors and fibronectin deposition. These results support and extend in vitro studies demonstrating that PAK contributes to activation of inflammatory pathways in endothelial cells by fluid shear stress. 相似文献