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1.
A-Rum Shin Hosung Sohn Choul Jae Won Byungsoo Lee Woo Sik Kim Hyun Bae Kang Hwa-Jung Kim Sang Nae Cho Sung Jae Shin 《Journal of microbiology (Seoul, Korea)》2010,48(4):502-511
Mycobacterium massiliense is an emerging pathogen and very similar to Mycobacterium abscessus of rapidly growing mycobacteria in the phenotype and genotype. Pathogenic bacteria secrete a diversity of factors into extracellular
medium which contribute to the bacterial pathogenicity. In the present study, we performed the comparative proteome analysis
of culture filtrate proteins from a clinical isolate of M. massiliense and M. abscessus strains using two-dimensional gel electrophoresis and liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS).
Interestingly, 9 proteins of M. massiliense were distinctly expressed from those of M. abscessus. Bioinformatic analysis of the identified proteins revealed that 3 unique proteins corresponded to serine/arginine rich protein,
membrane protein from Streptomyces coelicolor, and one hypothetical protein from Corynebacterium efficiens YS-314, respectively. Culture filtrate proteins from M. massiliense induced the release of pro-inflammatory cytokines from macrophages in a dose-dependent manner but not that from M. abscessus. Taken together, the functional study on the identified proteins uniquely produced from M. massiliense may provide not only the clues for the different pathogensis, but also help develop the diagnostic tools for the differentiation
between two mycobacterial species. 相似文献
2.
David Jáuregui-Zúñiga Yolanda Ortega-Ortega Martha Pedraza-Escalona Juan Pablo Reyes-Grajeda María Isabel Ruiz Carmen Quinto 《Plant Molecular Biology Reporter》2016,34(5):961-969
Legumes are unique in their ability to establish symbiotic interactions with rhizobacteria, providing a source of assimilable nitrogen; this symbiosis is regulated by complex signaling process between the plant and the bacteria. The participation of specific protein kinases during the initial steps of the nodulation process has been established. However, their substrates or the signaling networks implicated are not fully understood. Herein, a phosphoproteomic analysis of Phaseolus vulgaris roots treated for 24 h with specific Nod factors was performed using an immobilized metal ion affinity chromatography enrichment and two-dimensional gel electrophoresis approach with mass spectrometry identification. A total of 33 protein spots showing more than 1.5-fold shift were identified (17 protein spots in which the relative abundance increased and 16 that decreased). The majority of the identified root phosphoproteins displaying an increased relative abundance are presumed to have functions related to the biosynthesis and folding of proteins, energy metabolism, or cytoskeleton rearrangements, which reflect the metabolic status of the roots as being part of the developmental processes leading to nodule initiation and the importance of cytoskeleton rearrangement in the P. vulgaris–rhizobia symbiosis. The proteins in which relative abundance decreased are associated with defense and oxido-reduction processes, which could indicate a suppression of plant defense responses during the establishment of the rhizobia–legume interaction and an increase of reactive oxygen species production. 相似文献
3.
Hisano H Kimoto Y Hayakawa H Takeichi J Domae T Hashimoto R Abe J Asano S Kanazawa A Shimamoto Y 《Plant cell reports》2004,22(12):910-918
We have developed a new procedure for Agrobacterium-mediated transformation of plants in the genus Beta using shoot-base as the material for Agrobacterium infection. The frequency of regeneration from shoot bases was analyzed in seven accessions of sugarbeet (Beta vulgaris) and two accessions of B. maritima to select materials suitable for obtaining transformed plants. The frequency of transformation of the chosen accessions using Agrobacterium strain LBA4404 and selection on 150-mg/l kanamycin was found to be higher than that in previously published methods. Genomic DNA analysis and -glucuronidase reporter assays showed that the transgene was inherited and expressed in subsequent generations. In our method, shoot bases are prepared by a simple procedure, and transformation does not involve the callus phase, thus minimizing the occurrence of somaclonal variations. 相似文献
4.
Ju-Hong Zhang Li-Wen Sun Lin-Lin Liu Jie Lian Shao-Li An Xu Wang Jing Zhang Jun-Ling Jin Shan-Yu Li Jing-Hui Xi 《Plant Molecular Biology Reporter》2010,28(2):324-333
In this study, comparative proteomics was used to investigate the interaction of Spodoptera exigua and Arabidopsis thaliana. By using 2-D electrophoresis of differentially expressed proteins, combined with high-throughput matrix-assisted laser desorption/ionization
time of flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF/TOF MS, the changes in the abundance of proteins induced by
insect feeding were studied in A. thaliana. More than 1,100 protein spots were reproducibly detected on each gel. The intensities of 30 protein spots in particular
changed significantly, showing differences in volume of at least twofold. Among these, 17 protein spots were upregulated,
and 13 were downregulated following an 8-h insect feeding period. Nineteen insect-feeding-responsive proteins were identified,
all of which were involved in metabolic regulation, binding functions or cofactor requirement of protein, cell rescue, and
defense and virulence, as assessed by Munich Information Center for Protein Sequences function category. About 50% of these
were involved in metabolism, including transketolase, S-adenosylmethionine synthase 3, 2,3-biphosphoglycerate-independent phosphoglycerate mutase, beta-ureidopropionase, GDP-d-mannose 3′,5′-epimerase, and fatty acid synthase. The identification of insect-feeding-responsive proteins on Arabidopsis provides not only new insights into insect stress but also a good start for further investigation of their functions. Understanding
how the plant responses to insects in the proteomic level will provide tools for a better management of insect pest in the
field. 相似文献
5.
Chaikeeratisak V Somboonwiwat K Wang HC Lo CF Tassanakajon A 《Molecular biology reports》2012,39(5):6367-6377
6.
Autolysis is an important physiological process found in fungal cultivation. However, there is hitherto no report on the autolysis
of Pleurotus tuber-regium. We have investigated the enzymes secreted by temperature-induced (40°C as treatment versus 10°C as control) autolysis of
the mycelium of P. tuber-regium grown in submerged cultivation. A comparison between the intracellular proteins (inside the mycelium) and the extracellular
proteins (in the culture medium) of the treatment and control by proteomic analysis involving 2D PAGE and MALDI–TOF–MS was
made. Twenty-two up-regulated protein spots were detected and eight proteins were identified. They included proteasome which
participates in the ubiquitin–proteasome pathway; β-1,3-glucanosyltransferase and tubulin which are involved in the renewal
and repair of cell wall; protease and endoglucanase which promote the natural degradation of cell wall and cytoplasm; 14-3-3
protein which takes part in cell signal transduction; and two putative proteins presumably relate to the autolysis process.
These identified proteins suggest partially the metabolic processes of the autolysis in the P. tuber-regium mycelium. 相似文献
7.
Fuhua Bian Caixia Zheng Funing Qu Xueqin Gong Cuirong You 《Plant Molecular Biology Reporter》2010,28(1):22-31
Cyclamen persicum Mill. is a widely grown ornamental species that is clonally propagated by somatic embryogenesis. To better understand the
biology of somatic embryo development in C. persicum, detailed proteomic (two-dimensional gel electrophoresis) and mass spectrometric analyses of somatic embryos at globular,
torpedo, and germinating stages of development, along with nonembryogenic callus and zygotic embryos, were conducted. Of ~460
proteins resolved in two-dimensional gels, 35 proteins were differentially expressed and could be reproducibly displayed across
an isoelectric focusing range of 5 to 8. Among those proteins, five were constitutively expressed, 13 were upregulated, nine
were downregulated, and eight were deemed as novel proteins during the torpedo stage. A total of 35 protein spots were analyzed
by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and only four proteins were
identified and these were available in public protein databases. The remaining protein spots were subsequently analyzed by
MALDI-TOF-TOF-MS, and six proteins were then identified. These findings suggested that specific proteins are involved in the
regulation of somatic embryogenesis. 相似文献
8.
Salah F. Abou-Elwafa Bianca Büttner Friedrich J. Kopisch-Obuch Christian Jung Andreas E. Müller 《Molecular breeding : new strategies in plant improvement》2012,29(4):989-998
Bolting tendency in the crop species Beta vulgaris, which includes sugar beet, is a complex trait governed by various environmental cues, including prolonged periods of cold
temperatures over winter (vernalization) and photoperiod, and multiple genetic factors. Two loci which promote bolting in
the absence of vernalization are known in beet, the major bolting locus B on chromosome II and the B2 locus on chromosome IX. Here, genetic linkage and quantitative trait locus analyses in two populations derived from a cross
between a biennial genotype, which was identified in a phenotypic screen for EMS-induced bolting mutants and requires vernalization
to bolt, and an annual wild beet accession revealed the presence of a novel major bolting locus B4 which is linked to the B locus but promotes annual bolting independently of B. The genetic distance between B and B4 on chromosome II is 11 cM. A sequence-based marker was identified which co-segregates with bolting behavior and co-localizes
with the B4 locus. 相似文献
9.
Muhammad Rizwan Ingrid Miller Fareeha Tasneem Josef Böhm Manfred Gemeiner Ebrahim Razzazi-Fazeli 《Mycotoxin Research》2010,26(3):171-180
Genome sequencing for many important fungi has begun during recent years; however, there is still some deficiency in proteome
profiling of aspergilli. To obtain a comprehensive overview of proteins and their expression, a proteomic approach based on
2D gel electrophoresis and MALDI-TOF/TOF mass spectrometry was used to investigate A. ochraceus. The cell walls of fungi are exceptionally resistant to destruction, therefore two lysis protocols were tested: (1) lysis
via manual grinding using liquid nitrogen, and (2) mechanical lysis via rapid agitation with glass beads using MagNalyser.
Mechanical grinding with mortar and pestle using liquid nitrogen was found to be a more efficient extraction method for our
purpose, resulting in extracts with higher protein content and a clear band pattern in SDS-PAGE. Two-dimensional electrophoresis
gave a complex spot pattern comprising proteins of a broad range of isoelectric points and molecular masses. The most abundant
spots were subjected to mass spectrometric analysis. We could identify 31 spots representing 26 proteins, most of them involved
in metabolic processes and response to stress. Seventeen spots were identified by de novo sequencing due to a lack of DNA
and protein database sequences of A. ochraceus. The proteins identified in our study have been reported for the first time in A. ochraceus and this represents the first proteomic approach with identification of major proteins, when the fungus was grown under submerged
culture. 相似文献
10.
Molecular genetic studies of sugar beet (Beta vulgaris L.) are reviewed as a basis for the development of genomics of this species. The methods used to study structural and functional
genomics are considered. The results and their application to increase the efficiency of sugar beet breeding are discussed. 相似文献
11.
Extracellular protein profiles were compared for broth-grown cultures of Burkholderia pseudomallei and its avirulent close relative Burkholderia thailandensis. A number of protein bands were present in the B. pseudomallei profile but absent or less abundant in the B. thailandensis profile. Four such prominent secreted proteins were identified by using N-terminal sequencing coupled to searches of the B. pseudomallei genome sequence database. The genes for two proteins with similarity to carbohydrate-binding proteins, and a further protein homologous to known bacterial collagenases, were present in both B. pseudomallei and B. thailandensis. The putative collagenase gene was cloned and expressed as a fusion protein in Escherichia coli. Cell lysates from Escherichia coli containing the recombinant protein exhibited detectable gelatinase and collagenase activities. 相似文献
12.
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14.
Single-cell Raman microspectroscopy has the potential to report on the whole-cell chemical composition of bacteria, reflecting
metabolic status as well as growth history. This potential has been demonstrated through the discriminant functional analysis
of Raman spectral profiles (RSP) obtained from the soil and plant-associated bacterium Pseudomonas fluorescens SBW25, grown in vitro using defined media, and in planta using 3-month-old sugar beets (Beta vulgaris var. Roberta). SBW25 in vitro RSP data showed significant variation between those cells grown on different amino acids, sugars, TCA cycle intermediates,
rich King's B, and culture media derived from the sugar beet phytosphere. Raman analysis was also able to follow the transition
of SBW25 starved of carbon over a period of days, and SBW25 in planta RSP data also showed variation with significant differences between bacteria recovered from soil and the rhizosphere. SBW25
whole-cell chemical composition, and therefore growth and metabolic history, could be interpreted by coanalyzing in vitro and in planta RSP data. SBW25 recovered from the phytosphere was found to be more similar to SBW25 grown in vitro on Fru or Asp, rather than on Glc or Arg, and quite dissimilar to that resulting from carbon starvation. This suggests that
SBW25 growth in the phytosphere is generally neither carbon-catabolite-repressed nor carbon-limited. These findings demonstrate
that the analysis of single-cell RSP can differentiate between isogenic populations of bacteria with different metabolic histories
or after recovery from different parts of their natural environment. In addition, Raman analysis is also capable of providing
biologically relevant biochemical inferences, which might then be tested to uncover the mechanistic basis (biochemical–metabolic–genetic)
differentiating bacteria growing in complex environments and exposed to different conditions. 相似文献
15.
Besma Sghaier-Hammami Inmaculada Redondo-López Ana M. Maldonado-Alconada Sira Echevarría-Zomeño Jesús V. Jorrín-Novo 《Acta Physiologiae Plantarum》2012,34(3):905-922
The present work is directed at studying changes at the proteome level in Arabidopsis thaliana leaves in response to Pseudomonas syringae virulent (Pst) and avirulent (Pst avrRpt2) strains. Arabidopsis leaves were sampled from challenged plants at 4, 8 and 24 h post inoculation. Proteins were TCA–acetone–phenol
extracted and subjected to 2-DE (5–8 pH range) and MS/MS (MALDI–TOF–TOF) analysis. Out of 800 matched spots on each of the
36 gels analysed, 147 spots were either absent in at least one of the conditions studied (time or treatments; qualitative
variable spots) or differentially accumulated between time and treatments (quantitative variable spots). Out of the 24 proteins
successfully identified over TAIR10 database, 23 have not been reported previously in similar proteomics studies of the Arabidopsis thaliana–Pseudomonas syringae interaction. The exhaustive statistical analysis performed, including principal component and heat map, showed that 24 h
post inoculation can clearly discriminate the challenged plants from the control. The protein change occurred early (4 h post
inoculation) following the virulent pathogen infection, whereas the change occurred later (24 h post inoculation) following
the avirulent pathogen inoculation. Concerning the variable proteins, three behavioural groups can be observed: group 1 (common
protein changes in response to virulent and avirulent pathogen infection), group 2 (protein changes in response to virulent
pathogen infection) and group 3 (protein changes in response to avirulent pathogen infection). Differential identified proteins
following the pathogen infection belonged to different groups including those of oxidative stress defence, enzymes of metabolic
pathways and molecular chaperones. 相似文献
16.
Background
Little is known regarding the extent or targets of phosphorylation in mycoplasmas, yet in many other bacterial species phosphorylation is known to play an important role in signaling and regulation of cellular processes. To determine the prevalence of phosphorylation in mycoplasmas, we examined the CHAPS-soluble protein fractions of Mycoplasma genitalium and Mycoplasma pneumoniae by two-dimensional gel electrophoresis (2-DE), using a combination of Pro-Q Diamond phosphoprotein stain and 33P labeling. Protein spots that were positive for phosphorylation were identified by peptide mass fingerprinting using MALDI-TOF-TOF mass spectrometry. 相似文献17.
Proteomic analysis of somatic embryogenesis in <Emphasis Type="Italic">Vitis vinifera</Emphasis> 总被引:1,自引:0,他引:1
Two dimensional gel electrophoresis coupled to mass spectrometry has been used to study the somatic embryogenesis in Vitis vinifera, by comparing embryogenic and non embryogenic calluses of the Thompson seedless cv. More than 1,000 spots were reproducibly resolved in colloidal Coomassie brilliant blue stained gels over a pI nonlinear range of 3–10 in the first dimension and using homogeneous 12.5% polyacrylamide gels in the second dimension. The
expression pattern of 35 spots differed significantly between the two samples. These spots were processed by mass spectrometry
analysis and the protein identity was assigned by using both the non-redundant protein and EST databases. Several responsive
proteins, some already known to be involved in the somatic embryogenesis process while others, for the first time put into
relation with this process, have been described. Moreover, they have been subdivided in functional categories, and their putative
role is discussed in terms of their relevance in the somatic embryogenesis process. 相似文献
18.
The magnetotactic bacterium Magnetospirillum magnetotacticum MS-1 mineralizes the magnetite (Fe3O4) crystal and organizes a highly ordered intracellular structure, called the magnetosome. However, the iron transport system,
which supports the biogenesis of magnetite, is not fully understood. In this study, we first identified the expressions of
both the ferric and the ferrous iron transporter proteins in M. magnetotacticum. The cellular protein compositions of ferric and ferrous iron-rich cultures were examined using two-dimensional electrophoresis.
According to the gel patterns, two outer-membrane ferric-siderophore receptor homologues were identified as proteins strongly
induced in the ferrous iron-rich condition. Also, we identified for the first time that the ferrous iron transport protein,
FeoB, is expressed in the M. magnetotacticum cytoplasmic membrane using immunoblotting. 相似文献
19.
A two-parameter statistical model was used to predict the solubility of 96 putative virulence-associated proteins of Flavobacterium psychrophilum (CSF259-93) upon over expression in Escherichia coli. This analysis indicated that 88.5% of the F. psychrophilum proteins would be expressed as insoluble aggregates (inclusion bodies). These solubility predictions were verified experimentally
by colony filtration blot for six different F. psychrophilum proteins. A comprehensive analysis of codon usage identified over a dozen codons that are used frequently in F. psychrophilum, but that are rarely used in E. coli. Expression of F. psychrophilum proteins in E. coli was often associated with production of minor molecular weight products, presumably because of the codon usage bias between
these two organisms. Expression of recombinant protein in the presence of rare tRNA genes resulted in marginal improvements
in the expressed products. Consequently, Vibrio parahaemolyticus was developed as an alternative expression host because its codon usage is similar to F. psychrophilum. A full-length recombinant F. psychrophilum hemolysin was successfully expressed and purified from V. parahaemolyticus in soluble form, whereas this protein was insoluble upon expression in E. coli. We show that V. parahaemolyticus can be used as an alternate heterologous expression system that can remedy challenges associated with expression and production
of F. psychrophilum recombinant proteins. 相似文献
20.
Insect-pathogenic fungi penetrate their hosts directly through the cuticle. To better understand this process, we identified
genes that were up-regulated by Metarhizium anisopliae germinating and differentiating on Locusta migratoria wings using suppression subtractive hybridization (SSH). A total of 78 unique expressed sequence tags (ESTs) up-regulated
more than twofold during fungal growth on locust wings were identified. Among these 78 ESTs, 30 (38.5%) shared significant
similarity with NCBI annotated hypothetical proteins, 16 (20.5%) shared low similarity to known or predicted genes, might
represent novel genes, and 32 (41.0%) shared significant similarity with known proteins that are involved in various cell
and molecular processes such as cell metabolism, protein metabolism, stress response and defense, and cell structure and function.
Semi-quantitative RT-PCR analysis of six randomly selected genes confirmed the SSH results, verifying the fidelity of the
SSH data. The results of this study provide novel information on genes expressed during early stages of infection with M. anisopliae, and improve current understanding of fungal pathogenesis. 相似文献