• 제목/요약/키워드: Protein Expression Pattern

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Root Colonization and ISR-mediated Anthracnose Disease Control in Cucumber by Strain Enterobacter asburiae B1

  • Bharathkumar, S.;Park, Jin-Woo;Han, Ji-Hee;Park, Kyung-Seok
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.333-343
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    • 2009
  • Here, we show that an endophytic bacterial strain, Enterobacter asburiae B1 exhibits the ability to elicit ISR in cucumber, tobacco and Arabidopsis thaliana. This indicates that strain B1 has a widespread ability to elicit ISR on various host plants. In this study, E. asburiae strain B1 did not show antifungal activity against tested major fungal pathogens, Colletotrichum orbiculare, Botrytis cinerea, Phytophthora capsici, Rhizoctonia solani, and Fusarium oxysporum. Moreover, the siderophore production by E. asburiae strain B1 was observed under in vitro condition. In greenhouse experiments, the root treatment of strain B1 significantly reduced disease severity of cucumber anthracnose caused by fungal pathogen C. orbiculare compared to nontreated control plants. By root treatment of strain B1 more than 50% disease control against anthracnose on cucumber was observed in all greenhouse experiments. Simultaneously, under the greenhouse condition, the soil drench of strain B1 and a chemical inducer benzothiadiazole (BTH) to tobacco plants induced GUS activity which is linked with activation of PR promoter gene. Furthermore, in Arabidopsis thaliana plants the soil drench of strain B1 induced the defense gene expression of PR1 and PDF1.2 related to salicylic acid and jasmonic acid/ethylene signaling pathways, respectively. In this study, for the main focus on root colonization by strain B1 associated with defense responses, bacterial cells of strain B1 was tagged with the gfp gene encoding the green fluorescent protein in order to determine the colonization pattern of strain B1 in cucumber. The gfp-tagged B1 cells were found on root surface and internal colonization in root, stem, and leaf. In addition to this, the scanning electron microscopy observation showed that E. asburiae strain B1 was able to colonized cucumber root surface.

Effect of Grape Seed Proanthocyanidins on Tumor Vasculogenic Mimicry in Human Triple-negative Breast Cancer Cells

  • Luan, Yun-Yan;Liu, Zi-Min;Zhong, Jin-Yi;Yao, Ru-Yong;Yu, Hong-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.531-535
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    • 2015
  • Vasculogenic mimicry (VM) refers to the unique ability of highly aggressive tumor cells to mimic the pattern of embryonic vasculogenesis, which was associated with invasion and metastasis. The grape seed proanthocyanidins (GSPs) had attracted much attention as a potential bioactive anti-carcinogenic agent. However, GSPs regulation of VM and its possible mechanisms in a triple-negative breast cancer cells (TNBCs) remain not clear. Therefore, we examined the effect of GSPs on VM information in HCC1937 cell model. In this study, we identified the VM structure via the three-dimensional (3D) matrix in vitro. Cell viability was measured using the CCK8 assay. The effects of GSPs on human triple-negative breast cancer cells (TNBCs) HCC1937 in terms of related proteins of VM information were determined using western blot analysis. In vitro, the tubular networks were found in highly invasive HCC1937 cells but not in the non-invasive MCF-7 cells when plated on matrigel. The number of vascular channels was significantly reduced when cells were exposed in GSPs ($100{\mu}g$/ml) and GSPs ($200{\mu}g/mL$) groups (all p<0.001). Furthermore, we found that treatment with GSPs promoted transition of the mesenchymal state to the epithelial state in HCC1937 cells as well as reducing the expression of Twist1 protein, a master EMT regulator.GSPs has the ability to inhibit VM information by the suppression of Twist1 protein that could be related to the reversal of epithelial-to-mesenchymal (EMT) process. It is firstly concluded that GSPs may be an p otential anti-VM botanical agent for human TNBCs.

2-DE and MALDI-TOF MS-based identification of bovine whey proteins in milk collected soon after parturition

  • Lee, Jae Eun;Lin, Tao;Kang, Jung Won;Shin, Hyun Young;Lee, Joo Bin;Jin, Dong Il
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.635-643
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    • 2018
  • Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

Codon usage analysis of rice prolamine genes (쌀 저장 단백질 프롤라민 유전자 암호 분석)

  • Lee, Tae-Ho;Kim, Ju-Kon;Nahm, Baek-Hie
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.525-532
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    • 1993
  • To characterize the prolamines in rice cultivars, the complete coding sequences of 17 prolamine genes from the database were analyzed. According to the phylogenic analysis of the sequences, these genes could be classified into 4 groups, Group I to IV. The multiple alignment of the deduced amino acid sequences revealed that the four groups differ from one another in chain length caused by deletion of short internal amino acids or carboxyl terminal fragments. Each group was also found to have different amino acid composition with 1, 4, 10 and 30% of sulfur containing amino acids (methionine and cysteine) in Group I to IV prolamines, respectively. Also the isoelectric points of these groups showed the different values of 9.2, 8.2, 6.7 and 7.4. Finally, from the analysis of codon usage pattern of prolamine genes, the codon usage for arginine, serine, threonine, isoleucine, asparagine, aspartic acid, glutamic acid and cysteine were higly biased. In the analysis of the codon usage pattern, the relation of the fraction of G/C ending codons to effective codon numbers suggests the different translational efficiency in the expression of the prolamine multigenes.

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The virulence of Brucella abortus isolated from cattle in Korea (국내 소에서 분리한 Brucella abortus의 병원성 분석)

  • Lim, Jeong Ju;Kim, Jeong-Hwa;Kim, Dong Hyeok;Lee, Jin Ju;Kim, Dae Geun;Jun, Moo-Hyung;Kim, Sang Hun;Chang, Hong Hee;Lee, Hu Jang;Min, Won-Gi;Kim, Suk
    • Korean Journal of Veterinary Research
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    • v.51 no.1
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    • pp.15-20
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    • 2011
  • In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

Temporal Pattern of cAMP Concentrations and α-Actin mRNA Expression in Skeletal Muscle of Cimaterol-Fed Rats

  • Kim, Y.S.;Duguies, M.V.;Kim, Y.H.;Vincent, D.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.5
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    • pp.528-533
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    • 1997
  • Twenty four female Sprague-Dawley rats weighing about 190 g were used to examine changes in muscle cAMP concentrations and steady-state levels of skeletal muscle ${\alpha}$-actin mRNA during chronic administration of cimaterol, a ${\beta}$-adrenergic agonist. Cimaterol was mixed in a powdered rat diet at 10 mg/kg diet. At 3 and 21 days after the start of treatment, skeletal muscle and heart samples were collected for the measurement of cAMP concentrations and skeletal muscle ${\alpha}$-actin mRNA levels. Cimaterol increased (p < 0.01) body weight gain gradually during the first seven days of the trial period, but not thereafter. Most skeletal muscle weights and the ratio of muscle weight to body weight were increased (p < 0.05) by cimaterol treatment both at 3 and 21 days. Heart weight was also increased (p < 0.05) by cimaterol treatment at 3 and 21 days, but the ratio of heart weight to body weight was increased (p < 0.05) only at 3 day. Cimaterol decreased (p < 0.05) cAMP concentration of gastrocnemius muscle at both 3 and 21 days after treatment. However, cimaterol tended (p = 0.07) to increase cAMP concentration at 3 days in the heart. Cimaterol tended (p = 0.08) to increase the steady-state level of ${\alpha}$-actin mRNA by 60% in gastrocnemius muscle at 3 days but had no effect at 21 days. The results indicate that the pattern of hypertrophic response to chronic dietary administration of cimaterol is different between cardiac and skeletal muscle. In skeletal muscles it appears that the hypertrophy induced by cimaterol is partly due to stimulated myofibrillar protein synthesis at a pre-translational level.

Divergence of Genes Encoding Non-specific Lipid Transfer Proteins in the Poaceae Family

  • Jang, Cheol Seong;Jung, Jae Hyeong;Yim, Won Cheol;Lee, Byung-Moo;Seo, Yong Weon;Kim, Wook
    • Molecules and Cells
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    • v.24 no.2
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    • pp.215-223
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    • 2007
  • The genes encoding non-specific lipid transfer proteins (nsLTPs), members of a small multigene family, show a complex pattern of expressional regulation, suggesting that some diversification may have resulted from changes in their expression after duplication. In this study, the evolution of nsLTP genes within the Poaceae family was characterized via a survey of the pseudogenes and unigenes encoding the nsLTP in rice pseudomolecules and the NCBI unigene database. nsLTP-rich regions were detected in the distal portions of rice chromosomes 11 and 12; these may have resulted from the most recent large segmental duplication in the rice genome. Two independent tandem duplications were shown to occur within the nsLTP-rich regions of rice. The genomic distribution of the nsLTP genes in the rice genome differs from that in wheat. This may be attributed to gene migration, chromosomal rearrangement, and/or differential gene loss. The genomic distribution pattern of nsLTP genes in the Poaceae family points to the existence of some differences among cereal nsLTP genes, all of which diverged from an ancient gene. The unigenes encoding nsLTPs in each cereal species are clustered into five groups. The somewhat different distribution of nsLTP-encoding EST clones between the groups across cereal species imply that independent duplication(s) followed by subfunctionalization (and/or neofunctionalization) of the nsLTP gene family in each species occurred during speciation.

Expression of Caspase 3, Survivin, and p53 Protein in Urethane Induced Mouse Lung Carcinogenesis (Urethane으로 유발된 생쥐 폐샘암종 발생과정에서 Caspase 3, Survivin과 p53 단백 발현)

  • Shin, Jong Wook;Lee, Soo Hwan;Park, Eon Sub
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.251-260
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    • 2007
  • Purpose: An imbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis. Capase 3, survivin and p53 have been identified as important members of the apoptotic related proteins. This study evaluated the proliferating cell nuclear antigen(PCNA), apoptosis, apoptotic related protein such as capase 3, survivin and p53 using urethane-induced mouse lung carcinogenesis, which provides reproducible steps from hyperplasia to adenocarcinoma. Methods: Urethane was administered to the ICR mice through an intra-peritoneal injection, The mice were sacrificed at 5, 15, and 25 weeks after urethane intervention. The sequential morphological changes and immunohistochemical expression of PCNA, apoptosis, capase 3, survivin, and p53 were examined during mouse lung carcinogenesis. Results: During carcinogenesis, the sequential histological changes were observed from hyperplasia of type II pneumocytes, to anadenoma, and ultimately to an overt adenocarcinoma. The PCNA Labeling index (LI) was 9.6% in hyperplasia, 23.2% in adenoma, and 55.7% in adenocarcinoma, respectively. The apoptotic LI was 0.24% in hyperplasia, 1.25% in adenoma, and 5.27% in adenocarcinoma. A good correlation was observed between the PCNA LI and apoptotic LI. The expression of caspase 3 was remarkable- i.e., 46.7% in adenocarcinoma, in contrast to 15% in hyperplasia and 16% in adenoma. Survivin was detected weakly in the alveolar hyperplasia and showed an increasing expressional pattern in adenoma and adenocarcinoma. p53 expression was detected only in the adenocarcinoma lesions with an expression rate of 13.3%. The level of caspase 3 expression correlated with the increase in the apoptotic index. The positive expression of caspase 3 was associated with an increased apoptotic index. Conclusions: These results suggest that the PCNA LI and apoptotic LI might be useful markers for evaluating the risk of a malignant transformation. In addition, caspase, survivin and p53 might play a role in the early and late steges of urethane-induced mouse lung carcinogenesis.

Interleukin-8 gene expression in the human colon epithelial cell line, HT-29, exposed to Entamoeba histolytica (이질아메바에 의한 인체 대장상피세포주 HT-29에서의 interleukin-8 유전자의 발현)

  • 김정목;정현채
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.357-364
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    • 1995
  • The protozoan parasite, Entcmoeba histoIWticc, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to 5. histoIHtica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the (jading that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to 5. histolvtica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to 5. histoLvtica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polprnerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 H 107 molecules/ug of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA) . Lysates of 5. histoIVtica also induced expression of mRNA for IL-8 in colon epithelial cells. These results sugf:esc that acute inflammatory reaction by 5. histoIVticc may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

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Sequence and Expression Analysis of c-fos Proto-oncogene in Korean Cattle (HANWOO) (한우 c-fos 유전자의 염기서열 및 발현분석)

  • Yu, S.L.;Chung, H.J.;Jung, K.C.;Lee, J.H.;Cho, K.W.;Choi, J.G.;Na, K.J.;Sang, B.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.891-900
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    • 2003
  • Cellular FOS(c-fos) protein is a transcription factor that forms heterodimers mostly with c-jun family and stimulates the transcription of genes containing AP-1 regulatory elements. This c-fos expression can control growth and differentiation of various precursor cells including myoblasts. The controls by c-fos gene have been identified for affecting skeletal muscle fiber traits which are the key determinants of meat quality in pigs. As a first step for identifying the relationship between c-fos gene and meat quality traits in cattle, we fully sequenced 1,443 bp of Hanwoo c-fos mRNA and analyzed expression patterns from various organs and muscle tissues. The sequence identities of Hanwoo c-fos with that of human, pig and mouse showed 89.8%, 93.3% and 87%, respectively. Analyses of the northern blot showed high c-fos expressions were obtained in spleen and rib muscle from 7 organs and 9 different parts of muscles investigated. These results presented here can be used as a valuable marker for meat quality related traits in cattle with further verification.