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1.
Streptococcus suis is an important pathogen of pigs. In China, in addition to S. suis serotype 2, S. suis serotype 9 (SS9) is also a prevalent serotype. There is no vaccine available for SS9. An immunoproteome-based approach was
developed to identify SS9 immunogenic proteins for vaccine development. Secreted proteins extracted from SS9 strain GZ0565
were screened by two-dimensional Western blotting using convalescent sera from pigs. Protein spots were excised from preparative
gels and were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, which led to the
identification of ten immunogenic proteins (sortases, ABC transporter substrate-binding protein–maltose/maltodextrin, ABC
transporter periplasmic protein, CHAP domain containing protein, peptidoglycan-binding LysM, elongation factor Tu, elongation
factor G, thymidine kinase, molecular chaperone DnaK, hypothetical protein SSU98_2184). These novel immunogenic proteins,
which are encoded by genes that are reasonably conserved among SS9 strains, may be developed as antigens for further study
of SS9 vaccine. 相似文献
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Zhao Y Wang H Hammond RW Jomantiene R Liu Q Lin S Roe BA Davis RE 《Molecular genetics and genomics : MGG》2004,271(3):325-338
Spiroplasma kunkelii is a cell wall-free, helical, and motile mycoplasma-like organism that causes corn stunt disease in maize. The bacterium has a compact genome with a gene set approaching the minimal complement necessary for cellular life and pathogenesis. A set of 21 ATP-binding cassette (ABC) domains was identified during the annotation of a draft S. kunkelii genome sequence. These 21 ABC domains are present in 18 predicted proteins, and are components of 16 functional systems, which account for 5% of the protein coding capacity of the S. kunkelii genome. Of the 16 systems, 11 are membrane-bound transporters, and two are cytosolic systems involved in DNA repair and the oxidative stress response; the genes for the remaining three hypothetical systems harbor nonsense and/or frameshift mutations, so their functional status is doubtful. Assembly of the 11 multicomponent transporters, and comparisons with other known systems permitted functional predictions for the S. kunkelii ABC transporter systems. These transporters convey a wide variety of substrates, and are critical for nutrient uptake, multidrug resistance, and perhaps virulence. Our findings provide a framework for functional characterization of the ABC systems in S. kunkelii.Communicated by W. Goebel 相似文献
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Tetracycline-induced proteome of Pseudomonas putida KT2440 was analyzed by 2-D gel electrophoresis and matrix-assisted laser desorption ionization–time of flight/mass spectrum
(NALDI-TOF/MS) in order to understand cellular response to tetracycline. Of the proteins upregulated in a culture medium containing
subinhibitory concentration of tetracycline (50 μg/mL), we identified 38 proteins from cytosol and precipitated fractions
by peptide mass fingerprinting and mass spectrum/mass spectrum analysis. Various amino acids ABC transporters, a ribose ABC
transporter, and a sulfate ABC transporter were found to be upregulated. Protein synthesis-related proteins, stress proteins,
energy metabolic enzymes, and unknown proteins were also strongly induced. Of the identified upregulated proteins, several
proteins (isocitrate lyase, branched-chain amino acid ABC transporter, superoxide dismutase, etc.) were also upregulated under
phenol-induced stress condition. These results demonstrate that tetracycline at a high concentration induced comprehensive
stress in P. putida KT2440 and the global induction of proteins related to bacteria survival. Proteome analysis was found to be a useful tool
for the elucidation of antibiotic-induced proteins in the present study. 相似文献
6.
Hippophae rhamnoides is a hardy shrub capable of growing under extreme environmental conditions namely, high salt, drought and cold. Its ability to grow under extreme conditions and its wide application in pharmaceutical and nutraceutical industry calls for its in-depth analysis. N-glycoproteome mining by con A affinity chromatography from seedling was attempted. The glycoproteome was resolved on first and second dimension gel electrophoresis. A total of 48 spots were detected and 10 non-redundant proteins were identified by MALDI–TOF/TOF. Arabidopsis thaliana protein disulfide isomerase-like 1-4 (ATPDIL1-4) electron transporter, protein disulphide isomerase, calreticulin 1 (CRT1), glycosyl hydrolase family 38 (GH 38) protein, phantastica, maturase k, Arabidopsis trithorax related protein 6 (ATXR 6), cysteine protease inhibitor were identified out of which ATXR 6, phantastica and putative ATPDIL1-4 electron transporter are novel glycoproteins. Calcium binding protein CRT1 was validated for its calcium binding by stains all staining. GO analysis showed involvement of GH 38 and ATXR 6 in glycan and lysine degradation pathways. This is to our knowledge the first report of glycoproteome analysis for any Elaeagnaceae member. 相似文献
7.
Summary. Phosphate transport in bacteria occurs via a phosphate specific transporter system (PSTS) that belongs to the ABC family of
transporters, a multisubunit system, containing an alkaline phosphatase. DING proteins were characterized due to the N-terminal
amino acid sequence DINGG GATL, which is highly conserved in animal and plant isolates, but more variable in microbes. Most
prokaryotic homologues of the DING proteins often have some structural homology to phosphatases or periplasmic phosphate-binding
proteins. In E. coli, the product of the inducible gene DinG, possesses ATP hydrolyzing helicase enzymic activity. An alkaline phosphorolytic enzyme of the PSTS system was purified
to homogeneity from the thermophilic bacterium Thermus thermophilus. N-terminal sequence analysis of this protein revealed the same high degree of similarity to DING proteins especially to
the human synovial stimulatory protein P205, the steroidogenesis-inducing protein and to the phosphate ABC transporter, periplasmic
phosphate-binding protein, putative (P. fluorescens Pf-5). The enzyme had a molecular mass of 40 kDa on SDS/PAGE, exhibiting optimal phosphatase activity at pH 12.3 and 70 °C.
The enzyme possessed characteristics of a DING protein, such as ATPase, ds endonuclease and 3′ phosphodiesterase (3′-exonuclease)
activities and binding to linear dsDNA, displaying helicase activity on supercoiled DNA. Purification and biochemical characterization
of a T. thermophilus DING protein was achieved. The biochemical properties, N-terminal sequence similarities of this protein implied that the
enzyme belongs to the PSTS family and might be involved in the DNA repair mechanism of this microorganism.
Authors’ address: Assist. Prof. A. A. Pantazaki, Laboratory of Biochemistry, Department of Chemistry, Aristotle University
of Thessaloniki, Thessaloniki 54124, Greece 相似文献
8.
Seung II Kim Soo-Jung Kim Myung Hee Nam Soohyun Kim Kwon-Soo Ha Kye-Heon Oh Jong-Shin Yoo Young-Mok Park 《Current microbiology》2002,44(1):61-66
Acinetobacter lwoffii K24 is a soil bacterium that can use aniline as a sole carbon and nitrogen source (by β-ketoadipate pathway genes (cat genes)) and has two copies of catABC gene separately located on the chromosome. In order to identify aniline-induced proteins, two-dimensional electrophoresis
(2-DE) was applied to soluble protein fractions of A. lwoffii K24 cultured in aniline and succinate media. In the range of pH3–10, more than 370 spots were detected on the silver stained
gels. Interestingly, more than 20 spots were selectively induced on aniline-cultured bacteria. Twenty-three protein spots
of A. lwoffii K24 were analyzed by N-terminal microsequencing and internal microsequencing with in-gel digestion. Of 20 aniline induced
protein spots, we identified six β-ketoadipate pathway genes, one subunit of amino group transfer (putative subunit of aniline
oxygenase), malate dehydrogenase, putative ABC transporter, putative hydrolase, HHDD isomerase, and five unknown proteins.
Especially in case of two catechol 1,2-dioxygenases (CDI1 and CDI2), more than three isotypes were detected on the 2D gel. This study showed that the proteome analysis of A. lwoffii K24 may be helpful for identification of genes induced by aniline and understanding of their function in the cell.
Received: 2 April 2001 / Accepted: 14 May 2001 相似文献
9.
Polona Jamnik Maja Meglen Peter Raspor Nataša Poklar Ulrih 《World journal of microbiology & biotechnology》2010,26(9):1579-1586
Proteins from the extracellular medium of Aeropyrum pernix K1 were separated by two-dimensional electrophoresis and identified using mass spectrometry. Six different substrate-binding
proteins (SBPs) from the ATP-binding cassette (ABC) transporter family were identified: (1) ABC transporter SBP (Q9YC61);
(2) Branched-chain amino-acid ABC transporter, branched-chain amino-acid-binding protein (Q9YDJ6); (3) Oligopeptide ABC transporter,
oligopeptide-binding protein (Q9YBL5); (4) Probable ABC transporter SBP (Q9Y9N4); (5) ABC transporter SBP (Q9YBG7); (6) ABC
transporter SBP (Q9YFD7). Based on their orthology, division into the following classes was predicted: (1) multiple sugar-transport
system SBPs; (2) peptide/nickel-transport system SBPs; and (3) branched-chain amino-acid-transport system SBPs. Further bioinformatic
analyses showed that the identified SBPs differ in motif and in transmembrane-domain and signal-peptide organisation. Additionally,
for all of these SBPs, sequence homology was found for archaeal proteins, and homologous proteins in bacteria were also found
for the ABC transporter SBP Q9YBG7 and the ABC transporter SBP Q9YFD7. This is the first study, where different ABC SBPs from
the extracellular medium of A. pernix have been identified using the combined methodology of two-dimensional electrophoresis and mass spectrometry. 相似文献
10.
Plant aquaporins are believed to facilitate water transport across cell membranes. However, the relationship between aquaporins
and drought resistance in plants remains unclear. VfPIP1, a putative aquaporin gene, was isolated from Vicia faba leaf epidermis, and its expression was induced by abscisic acid (ABA). Our results indicated that the VfPIP1 protein was
localized in the plasma membrane, and its expression in V. faba was induced by 20% polyethylene glycol 6000. To further understand the function of VfPIP1, we obtained VfPIP1-expressing transgenic Arabidopsis thaliana plants under the control of the CaMV35S promoter. As compared to the wild-type control plants, the transgenic plants exhibited
a faster growth rate, a lower transpiration rate, and greater drought tolerance. In addition, the stomata of the transgenic
plants closed significantly faster than those of the control plants under ABA or dark treatment. These results suggest that
VfPIP1 expression may improve drought resistance of the transgenic plants by promoting stomatal closure under drought stress. 相似文献
11.
Lu Xianwen Sun Kun Ma Ruijun Zhang Hui Su Xue Wang Mingli 《Frontiers of Biology in China》2006,1(3):318-322
Behaviors of 18 species of birds eating fruits of Hippophae rhamnoides spp. sinensis were observed from September 2003 to March 2004. Their foraging patterns were found to be very different and can be divided
into five classes: (1) direct swallowing the fruits on crown of the shrubs and sometimes regurgitating seeds soon after; (2)
carrying the fruits to their perching sites and swallowing; (3) pecking the fruits from the shrubs to the ground, eating pulp
and seeds but leaving pericarp; (4) pecking through the pericarp, eating pulp and leaving pericarp and seeds; (5) pecking
through the pericarp on the top of fruits, and only eating seeds. These foraging patterns have different effects on seed dispersal
of H. rhamnoides spp. sinensis. The germination experiment of three groups of seeds (seeds from feces, dry fruits and extracted seeds from dry fruits) was
carried out. Although ingestion processes of birds had some adverse effects on the seed germination of H. rhamnoides spp. sinensis, the seeds from feces still have a relatively higher germination ratio. H. rhamnoides spp. sinensis provides food to a variety of frugivorous birds, and the birds disperse its seeds. Thus, a mutually beneficial relationship
between the bird and the seed is formed.
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Translated from Chinese Journal of Ecology, 2005, 24(6): 635–638 [译自: 生态学杂志, 2005, 24(6): 635–638] 相似文献
12.
We adapted PCR-based signature-tagged mutagenesis (STM) to Pseudomonas aeruginosa. A collection of 1056 mutants was screened in a chronic lung infection rat model. Thirteen mutants were confirmed to be attenuated. Analysis revealed that these STM mutants represented transposon insertions into eight genes previously described in databases, three genes encoding proteins sharing identity with hypothetical proteins and two genes that shared no significant identity with sequences in databases. Five strains mutated in genes involved in protein degradation, stress tolerance, cation transport, ABC transporter, and an unknown protein were shown to be highly attenuated when tested individually in the rat chronic lung infection model. 相似文献
13.
In this study, secreted Corynebacterium glutamicum proteins were investigated by two-dimensional gel electrophoresis. Around 100 spots observed in the pH range 4.5–5.5 had
molecular masses that varied from 10 to 50 kDa. Upon N-terminal amino acid sequence analysis by Edman degradation, two of
them were hits to two hypothetical proteins encoded by cgR_1176 and cgR_2070 on C. glutamicum R genome, respectively. Active-form α-amylase derived from Geobacillus stearothermophilus was successfully secreted by using the predicted cgR_1176 and cgR_2070 signal sequences, indicating that these hypothetical proteins were secreted proteins. Analysis using a disruption mutant
of the twin-arginine translocation (Tat) export pathway machinery of C. glutamicum suggested that one is Tat pathway dependent secretion while the other is independent of the pathway. Our results demonstrate
that C. glutamicum can secrete exoproteins by using its own signal sequences, indicating its potential as a host for protein productions. 相似文献
14.
Summary. Plants have evolved many mechanisms to cope with adverse environmental stresses. Abscisic acid (ABA) accumulates significantly
in plant cells in response to drought conditions, and this is believed to be a major mechanism through which plants enhance
drought tolerance. In this study, we explore the possible mechanisms of osmotic stress perception by plant cells and the consequent
induction of ABA biosynthesis. Immunoblotting and immunofluorescence localization experiments, using a polyclonal antibody
against human integrin β1, revealed the presence of a protein in Zea mays roots that is similar to the integrin proteins of animals and mainly localized in the plasma membrane. Treatment with GRGDS,
a synthetic pentapeptide containing an RGD domain, which interacted specifically with the integrin protein and thus blocked
the cell wall–plasma membrane interaction, significantly inhibited osmotic stress-induced ABA biosynthesis in cells, and the
GRGDS analog which does not contain the RGD domain had no effect. Our results show that a strong interaction exists between
the cell wall and plasma membrane and that this interaction is largely mediated by integrin-like proteins. They also imply
that the cell wall and/or cell wall–plasma membrane interaction plays important roles in the perception of osmotic stress.
Accordingly, we conclude that the cell wall and/or cell wall–plasma membrane interaction mediated by the integrin-like protein
plays important roles in osmotic stress-induced ABA biosynthesis in Zea mays.
Correspondence: J. S. Liang, College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, People’s Republic
of China. 相似文献
15.
Genetic analysis for drought resistance of rice at reproductive stage in field with different types of soil 总被引:9,自引:0,他引:9
Yue B Xiong L Xue W Xing Y Luo L Xu C 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,111(6):1127-1136
Drought resistance of rice is a complex trait and is mainly determined by mechanisms of drought avoidance and drought tolerance. The present study was conducted to characterize the genetic basis of drought resistance at reproductive stage in field by analyzing the QTLs for drought response index (DRI, normalized by potential yield and flowering time), relative yield, relative spikelet fertility, and four traits of plant water status and their relationships with root traits using a recombinant inbred population derived from a cross between an indica rice and upland rice. A total of 39 QTLs for these traits were detected with individual QTL explained 5.1–32.1% of phenotypic variation. Only two QTLs for plant water status were commonly detected in two environments, suggesting different mechanisms might exist in two types of soil conditions. DRI has no correlation with potential yield and flowering time under control, suggesting that it can be used as a good drought resistance index in field conditions. The co-location of QTLs for canopy temperature and delaying in flowering time suggested a usefulness of these two traits as indexes in drought resistance screening. Correlation and QTL congruence between root traits and putative drought tolerance traits revealed that drought avoidance (via thick and deep root traits) was the main genetic basis of drought resistance in sandy soil condition, while drought tolerance may play more role in the genetic basis of drought resistance in paddy soil condition. Therefore, both drought mechanisms and soil textures must be considered in the improvement of drought resistance at reproductive stage in rice. 相似文献
16.
【背景】解析植物微生物群落结构是利用微生物组工程强化植物抗生物和非生物胁迫水平、提高农林产品质量和品质的基础。固氮根瘤是沙棘具有抗旱、抗寒和抗贫瘠等多种优良生物性状的关键。【目的】比较分析沙棘根际土和根瘤内细菌群落结构的组成及影响因素,为揭示沙棘-弗兰克氏菌共生和植物-微生物互作协同抗逆机制提供理论基础。【方法】从辽宁、陕西和山西采集样品,通过16SrRNA基因V3–V4可变区的高通量测序技术,采用生物信息学方法比较分析沙棘根际土和根瘤内细菌群落组成和丰度差异,并探索土壤土理化性质对根际土细菌群落结构的影响。【结果】沙棘根际土和根瘤内的细菌群落均以放线菌门(Actinobacteria)和变形菌门(Proteobacteria)为主要优势菌门,且根瘤内弗兰克氏菌属(Frankia)为绝对优势菌属;根际土前10个优势菌门的丰度在三地样品间均存在显著差异,仅存在唯一共有的优势菌属(鞘氨醇单胞菌属,Sphingomonas),且前35个优势属中有27个属在三地间存在明显丰度差异;土壤pH和速效钾是沙棘根际土细菌群落多样性的主要影响因子;根瘤内优势门和属在三省份间存在高度的保守性,仅异根瘤菌属... 相似文献
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A ciprofloxacin-resistant mutant of Clostridium perfringens, strain VPI-C, which had stable mutations in the topoisomerase genes, accumulated less norfloxacin and ethidium bromide than the wild type, strain VPI. Efflux pump inhibitors both increased the accumulation of ethidium bromide by cells of the mutant and enhanced their sensitivity to this toxic dye. Cloning a gene, which codes for a putative ABC transporter protein (NP_562422) of 527 amino acids, from the mutant strain VPI-C into the wild-type strain VPI not only reduced the accumulation of ethidium bromide by the recombinant strain but also reduced its sensitivity to norfloxacin and ciprofloxacin. Efflux pump inhibitors decreased the rate at which ethidium bromide was removed from the cells of the recombinant strain. It appears that the putative ABC transporter protein (NP_562422) may contribute to extrusion of drugs from C. perfringens. 相似文献
19.
Caiyun He Jianguo Zhang Aiguo Duan Shuxing Zheng Honggang Sun Lihua Fu 《Trees - Structure and Function》2008,22(6):803-813
Proteomic analysis provides a powerful method of studying plant responses to stress at the protein level. In order to study
stress-responsive molecular mechanisms for Populus × euramericana cv. ‘74/76’, one of the most important forest plantation tree species in subtropical and temperate regions, we analyzed the
response of 2-year-old cuttings of P. × euramericana cv. ‘74/76’ to drought and high temperature using two-dimensional gel electrophoresis. More than 1,000 reproducible leaf
proteins were detected in the controls and treatments, and 26 proteins were found to change notably in abundance. We identified
13 proteins affected by drought stress and 11 proteins affected by high temperature. These proteins are mainly involved in
photosynthesis such as ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit and putative photosystem I reaction center
subunit II precursor, and detoxification (manganese superoxide dismutase and methionine sulfoxide reductase A). Furthermore,
the level of the photosynthesis proteins affected greatly by the imposed stress conditions was consistent with the observed
noticeable decrease in net photosynthesis rate. These studies provides a fundamental data for future research on responses
to drought and high temperature, two major factors limiting the growth of forest trees during summer under recent climatic
warming. 相似文献
20.
Copper (Cu) accumulation and tolerance mechanisms in Elsholtzia haichowensis, an indicator plant of Cu mines, were investigated under hydroponics supplied with different concentrations (0.32, 50.0,
100.0 and 200.0 μM) of Cu for 8 days. Cu at 100 and 200 μM significantly decreased the root dry weight, but had no significant
effect on shoot dry weight. The plants grown in the presence of 200 μM Cu accumulated 288 and 7626 μg g−1 DW total Cu in the shoots and roots, respectively. A greater proportion of accumulated Cu was water-soluble accounting for
42–93% of the total Cu content in the shoots. The concentrations of reduced glutathione (GSH) and protein thiols were significantly
enhanced under excess Cu supply. However, the concentrations of these compounds, particularly protein thiols, were much higher
in the leaves than that in the roots. Three UV-absorbing peaks could be eluted out through gel filtration chromatography on
Sephadex G-50. A large amount of Cu was detected in the UV-absorbing peaks in 40–50 and 70–90 ml elution fractions of the
root extract, and in 40–50 and 120–140 ml elution fractions of the leaf extract. The results suggested that the adaptive Cu
tolerance mechanism in E. haichowensis might involve the active participation of protein thiols which had a more important role in the leaves than in the roots. 相似文献