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Tracking elusive cargo: Illuminating spatio‐temporal Type 3 effector protein dynamics using reporters 下载免费PDF全文
Type 3 secretion systems form an integral part of the arsenal of many pathogenic bacteria. These injection machines, together with their cargo of subversive effector proteins, are capable of manipulating the cellular environment of the host in order to ensure persistence of the pathogen. In order to fully appreciate the functions of Type 3 effectors, it is necessary to gain spatio‐temporal knowledge of each effector during the process of infection. A number of genetic modifications have been exploited in order to reveal effector protein secretion, translocation and subsequent activity, and localisation within host cells. In this review, we will discuss the many available approaches for tracking effector protein dynamics and discuss the challenges faced to improve the current technologies and gain a clearer picture of effector protein function. 相似文献
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Gao Y Pagnon J Feng HC Konstantopolous N Jowett JB Walder K Collier GR 《Biochemical and biophysical research communications》2007,356(3):636-641
SEPS1 (also called selenoprotein S, SelS, Tanis or VIMP) is a selenoprotein, localized predominantly in the ER membrane and also on the cell surface. In this report, we demonstrate that SEPS1 protein is also secreted from hepatoma cells but not from five other types of cells examined. The secretion can be abolished by the ER-Golgi transport inhibitor Brefeldin A and by the protein synthesis inhibitor cycloheximide. Using a sandwich ELISA, SEPS1 was detected in the sera of 65 out of 209 human subjects (31.1%, average=15.7+/-1.1 ng/mL). Fractionation of human serum indicated that SEPS1 was associated with LDL and possibly with VLDL. The function of plasma SEPS1 is unclear but may be related to lipoprotein metabolism. 相似文献
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An E. coli expression system optimized for DELLA proteins 总被引:1,自引:0,他引:1
Sun X Frearson N Kirk C Jones WT Harvey D Rakonjac J Foster T Al-Samarrai T 《Protein expression and purification》2008,58(1):168-174
The DELLA proteins are involved in regulation of plant growth in response to phytohormonal signals such as GA, ethylene, and auxin. They have become one of most challenging and active area of research due to their fundamental roles in plant biology. Here, we describe the first successful expression of the N-terminal domains of DELLA proteins of Arabidopsis thaliana and Malus domestica in Escherichia coli system which will be used to produce monoclonal antibodies for profiling protein micro-arrays. Optimizations of the cloning, expression, and purification using specific tags have been discussed. 相似文献
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Karen A. Stockin Vicky Binedell † Nicky Wiseman ‡ Dianne H. Brunton § Mark B. Orams # 《Marine Mammal Science》2009,25(2):283-301
Here we present the first data describing the behavior of common dolphins ( Delphinus sp.) in the Hauraki Gulf, New Zealand. Activity budgets are used to assess the effects of diel, season, depth, sea surface temperature, group size, and composition on dolphin behavior. Additionally, the presence/absence of Bryde's whale ( Balaenoptera brydei ) and Australasian gannet ( Morus serrator ) is examined in relation to dolphin behavior. Behavioral data were collected from 686 independent dolphin groups during boat-based surveys conducted between February 2002 and January 2005. Foraging (46.7%) and social (7.2%) were the most and least frequently observed behaviors, respectively. Travel (28.9%), mill (9.5%), and rest (7.7%) accounted for the remainder of the activity budget. Behavior varied seasonally, with the highest proportion of foraging and resting groups observed during the spring and autumn, respectively. Behavior also varied with water depth, with foraging animals observed in the deepest and resting groups recorded in the shallowest regions of the Hauraki Gulf. A correlation between group size and behavior was evident, although behavior did not vary with the composition of dolphin groups. Resting, milling, and socializing animals were more frequently observed in smaller group sizes. Foraging behavior was prevalent in both small and large group sizes, suggesting foraging plasticity exists within this population. Behavior differed between single- and multispecies groups, with foraging more frequent in multispecies groups. Resting, milling, or socializing was rarely observed in the presence of any associated species, indicating the primary mechanism for association is likely prey related. 相似文献
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Christine Decaestecker Xavier Moles Lopez Nicky D'Haene Isabelle Roland Saad Guendouz Christophe Duponchelle Alix Berton Olivier Debeir Isabelle Salmon 《Proteomics》2009,9(19):4478-4494
Antibody‐based proteomics applied to tissue microarray (TMA) technology provides a very efficient means of visualizing and locating antigen expression in large collections of normal and pathological tissue samples. To characterize antigen expression on TMAs, the use of image analysis methods avoids the effects of human subjectivity evidenced in manual microscopical analysis. Thus, these methods have the potential to significantly enhance both precision and reproducibility. Although some commercial systems include tools for the quantitative evaluation of immunohistochemistry‐stained images, there exists no clear agreement on best practices to allow for correct and reproducible quantification results. Our study focuses on practical aspects regarding (i) image acquisition (ii) segmentation of staining and counterstaining areas and (iii) extraction of quantitative features. We illustrate our findings using a commercial system to quantify different immunohistochemistry markers targeting proteins with different expression patterns (cytoplasmic, nuclear or membranous) in colon cancer or brain tumor TMAs. Our investigations led us to identify several steps that we consider essential for standardizing computer‐assisted immunostaining quantification experiments. In addition, we propose a data normalization process based on reference materials to be able to compare measurements between studies involving different TMAs. In conclusion, we recommend certain critical prerequisites that commercial or in‐house systems should satisfy in order to permit valid immunostaining quantification. 相似文献
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Jessica K. Walker Nicky McGrath Danielle L. Nilsson Natalie K. Waran Clive J. C. Phillips 《Anthrozo?s》2014,27(2):251-266
Gender plays a significant role in influencing people's attitudes toward animals, however, little is known about how it influences their attribution of emotions to animals. To investigate the role that gender plays in public attitudes toward animals' experience of emotions and beliefs about whether animals can grieve, a face-to-face survey of 1,000 members of the general public was carried out in Brisbane, Australia. Potential respondents were asked to complete a 10-min “social attitudinal” survey. Males were significantly less likely than females to believe that animals experience complex emotions, including depression (p < 0.05), anxiety (p < 0.05), love (p < 0.01), and grief (p < 0.05), but did not differ in regard to basic emotions including distress, fear, happiness, anger, sadness, and fear. Males were also less likely to believe that animals show some behavioral (eating p < 0.05; vocalizing p < 0.01) changes when they experience grief (p < 0.05) and that animals grieve as a result of separation from a conspecific (p < 0.005). These results demonstrate a greater skepticism in males, compared with females, regarding the attribution of emotions to animals. 相似文献
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Anne L. Harrison Nicky Petkov Daniel Mitev Georgy Popgeorgiev Benedict Gove Geoff M. Hilton 《Biodiversity and Conservation》2018,27(1):167-188
The management of Arctic migrant geese is complex, because they frequently use landscapes under intensive human use, and are conflict species in multiple respects. Some populations are of high conservation concern, but they also cause agricultural damage, are quarry for hunters, and may be particularly sensitive to infrastructure developments. In Bulgarian Dobrudzha, large wintering populations of greater white-fronted geese Anser albifrons and red-breasted geese Branta ruficollis feed in agricultural land, and cause management dilemmas. We developed linear models to investigate fine- and meso-scale foraging habitat selection of geese foraging on winter wheat in the area, and used these models to make suggestions for zoning landscape use in order to reduce conflict and conserve geese. Habitat selection was scale-dependent. Geese selected fields that were near to major roosts and had low proximity to roads and tree-lines, which may be a proxy for hunting disturbance. We found some evidence for selection of wheat fields with high nutritional quality. Within fields, geese strongly avoided features which cause landscape ‘clutter’: power-lines, tree-lines and wind-turbines, but primarily over distances of less than a few hundred metres. Optimal management might involve encouraging goose populations to feed in areas close to roosts, by means of agri-environmental measures and creation of hunting-free refuges. This would allow efficient use of agri-environment funds, might reduce conflict with farmers, and would mean that infrastructure development—notably wind farms—could be sited at greater distance from roosts with relatively minor impact on foraging habitat availability. 相似文献