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141.
142.
Low-temperature increases the yield of biologically active herring antifreeze protein in Pichia pastoris 总被引:6,自引:0,他引:6
Li Z Xiong F Lin Q d'Anjou M Daugulis AJ Yang DS Hew CL 《Protein expression and purification》2001,21(3):438-445
Antifreeze proteins and antifreeze glycoproteins are structurally diverse molecules that share a common property in binding to ice crystals and inhibiting ice crystal growth. Type II fish antifreeze protein of Atlantic herring (Clupea harengus harengus) is unique in its requirement of Ca(2+) for antifreeze activity. In this study, we utilized the secretion vector pGAPZalpha A to express recombinant herring antifreeze protein (WT) and a fusion protein with a C-terminal six-histidine tag (WT-6H) in yeast Pichia pastoris wild-type strain X-33 or protease-deficient strain SMD1168H. Both recombinant proteins were secreted into the culture medium and properly folded and functioned as the native herring antifreeze protein. Furthermore, our studies demonstrated that expression at a lower temperature increased the yield of the recombinant protein dramatically, which might be due to the enhanced protein folding pathway, as well as increased cell viability at lower temperature. These data suggested that P. pastoris is a useful system for the production of soluble and biologically active herring antifreeze protein required for structural and functional studies. 相似文献
143.
Yeh S Moffatt BA Griffith M Xiong F Yang DS Wiseman SB Sarhan F Danyluk J Xue YQ Hew CL Doherty-Kirby A Lajoie G 《Plant physiology》2000,124(3):1251-1264
144.
Alexander Hew Dale Crooke 《Arts Education Policy Review》2019,120(1):19-32
This article explores the extent to which recent policy-based advocacy for music in Australian schools has been guided by economic arguments. This exploration was driven by emerging commentary on the rise of neo-social education policy, which is said to have reframed equality and social justice in education as tools for economic agendas. To explore the presence of this neo-social ideology in school music advocacy, key Australian policy documents have been analyzed using Critical Discourse Analysis. Results show that while current advocacy does champion social and subjective benefits, these benefits are ultimately justified in economistic terms. While caveats to these findings are discussed, the argument is made for new education policies that adequately value subjective wellbeing and social justice as goals in their own right. 相似文献
145.
Proteomic analysis of protein profiles during early development of the zebrafish, Danio rerio 总被引:1,自引:0,他引:1
In the present study, profiles of protein expression were examined during early development of zebrafish, an increasingly popular experimental model in vertebrate development and human diseases. By 2-DE, an initial increase in protein spots from 6 h post-fertilization (hpf) to 8-10 hpf was observed. There was no dramatic change in protein profiles up to 18 hpf, but significant changes occurred in subsequent stages. Interestingly, 49% of the proteins detected at 6 hpf remained detectable by 1 week of age. To map the protein expression patterns in 2-D gels, MALDI-TOF/TOF MS was employed to identify selected protein spots from early embryos. 108 protein spots were found to match known proteins and they were derived from 55 distinct genes. Interestingly, 11 (20%) of them produced multiple protein isoforms or distinct cleavage products. Although deyolked embryos were used in the analysis, a large number of vitellogenin derivatives remained prominently present in the embryos. Other than these, most of the identified proteins are cytosolic, cytoskeletal and nuclear proteins, which are involved in diversified functions such as metabolism, cytoskeleton, translation, protein degradation, etc. Some of the proteins with interesting temporal expression profiles during development are further discussed. 相似文献
146.
Pankaj K Bhavsar Bruce D Levy Mark J Hew Michael A Pfeffer Shamsah Kazani Elliot Israel Kian Fan Chung 《Respiratory research》2010,11(1):71
Background
An imbalance in the generation of pro-inflammatory leukotrienes, and counter-regulatory lipoxins is present in severe asthma. We measured leukotriene B4 (LTB4), and lipoxin A4 (LXA4) production by alveolar macrophages (AMs) and studied the impact of corticosteroids.Methods
AMs obtained by fiberoptic bronchoscopy from 14 non-asthmatics, 12 non-severe and 11 severe asthmatics were stimulated with lipopolysaccharide (LPS,10 μg/ml) with or without dexamethasone (10-6M). LTB4 and LXA4 were measured by enzyme immunoassay.Results
LXA4 biosynthesis was decreased from severe asthma AMs compared to non-severe (p < 0.05) and normal subjects (p < 0.001). LXA4 induced by LPS was highest in normal subjects and lowest in severe asthmatics (p < 0.01). Basal levels of LTB4 were decreased in severe asthmatics compared to normal subjects (p < 0.05), but not to non-severe asthma. LPS-induced LTB4 was increased in severe asthma compared to non-severe asthma (p < 0.05). Dexamethasone inhibited LPS-induced LTB4 and LXA4, with lesser suppression of LTB4 in severe asthma patients (p < 0.05). There was a significant correlation between LPS-induced LXA4 and FEV1 (% predicted) (rs = 0.60; p < 0.01).Conclusions
Decreased LXA4 and increased LTB4 generation plus impaired corticosteroid sensitivity of LPS-induced LTB4 but not of LXA4 support a role for AMs in establishing a pro-inflammatory balance in severe asthma. 相似文献147.
Tan HT Tan S Lin Q Lim TK Hew CL Chung MC 《Molecular & cellular proteomics : MCP》2008,7(6):1174-1185
Colorectal cancer is one of the most common cancers in developed countries, and its incidence is negatively associated with high dietary fiber intake. Butyrate, a short-chain fatty acid fermentation by-product of fiber induces cell maturation with the promotion of growth arrest, differentiation, and/or apoptosis of cancer cells. The stimulation of cell maturation by butyrate in colonic cancer cells follows a temporal progression from the early phase of growth arrest to the activation of apoptotic cascades. Previously we performed two-dimensional DIGE to identify differentially expressed proteins induced by 24-h butyrate treatment of HCT-116 colorectal cancer cells. Herein we used quantitative proteomics approaches using iTRAQ (isobaric tags for relative and absolute quantitation), a stable isotope labeling methodology that enables multiplexing of four samples, for a temporal study of HCT-116 cells treated with butyrate. In addition, cleavable ICAT, which selectively tags cysteine-containing proteins, was also used, and the results complemented those obtained from the iTRAQ strategy. Selected protein targets were validated by real time PCR and Western blotting. A model is proposed to illustrate our findings from this temporal analysis of the butyrate-responsive proteome that uncovered several integrated cellular processes and pathways involved in growth arrest, apoptosis, and metastasis. These signature clusters of butyrate-regulated pathways are potential targets for novel chemopreventive and therapeutic drugs for treatment of colorectal cancer. 相似文献
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150.
The normal supply of growth substances to a young soybean plant was altered by removing the plant's apical meristem and replacing this meristem with an aqueous solution of either indole-3-acetic acid (IAA), gibberellic acid (GA), or water. The length of each experiment was 1 hr. In the middle of it, 14CO2 was administered to one of the primary leaves of the plant, and at the end distribution of 14C in various parts of the plant was determined. It was found that an addition of growth substances stimulated translocation in three different ways. Both IAA and GA increased the total amounts of sucrose-14C translocated, increased the rate of its translocation, and affected the distribution pattern of translocated sucrose throughout the plant. Experiments using IAA-14C have shown that the action of IAA is on the longitudinal translocation in the stem and not on the transfer of photosynthate from the mesophyll to the conducting tissues of the leaf. 相似文献