• Title/Summary/Keyword: High-level expression

Search Result 1,204, Processing Time 0.028 seconds

Immunization of Mice with Recombinant Brucella abortus Organic Hydroperoxide Resistance (Ohr) Protein Protects Against a Virulent Brucella abortus 544 Infection

  • Hop, Huynh Tan;Reyes, Alisha Wehdnesday Bernardo;Simborio, Hannah Leah Tadeja;Arayan, Lauren Togonon;Min, Won Gi;Lee, Hu Jang;Lee, Jin Ju;Chang, Hong Hee;Kim, Suk
    • Journal of Microbiology and Biotechnology
    • /
    • v.26 no.1
    • /
    • pp.190-196
    • /
    • 2016
  • In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.

A Note on Teaching Method of Addition and Subtraction between Korea and New Zealand Primary School (한국과 뉴질랜드 초등학교 저학년의 덧셈과 뺄셈 지도방법에 관한 고찰)

  • Choi, Chang Woo
    • East Asian mathematical journal
    • /
    • v.31 no.4
    • /
    • pp.505-525
    • /
    • 2015
  • The purpose of this study is to analyze teaching method of addition and subtraction of whole number in Korea and New Zealand lower grade textbook and to get some suggestive points to develop mathematics curriculum and for a qualitative improvement of textbook. To do this, we will analyze focusing on teaching material, type and method of teaching, cases of real teaching and in the case of New Zealand, we will analyze portfolios together to see what kind of things do they deal with related to addition and subtraction. From these analyzing, the results are as follows: First, the guideline of accomplishment of group of year are stated in 2009 revised curriculum in Korea but it is rough. On the other hand, the level of accomplishment from kindergarten to high school are stated divided by eight kinds of thing in New Zealand curriculum. Second, there were common and different points in the aspect of teaching material. The common points are that both of our Korea and New Zealand are using materials related to real life intimately and the diifferent points are to use technology such as calculator and computer. They are more widely used in New Zealand than our Korea. Third, Korea had used routine method mainly but New Zealand had used method to develop creativity of learner such as to write problem corresponding to expression, posing problem corresponding to information, to complete table and find pattern and to write word problem to explain pattern and so on. Fourth, we could see special calculation strategies in the case of teaching addition and subtraction such as concept of double, compensation, various strategy based on counting of number, addition of the same number, magic square, near-double which are not finding in our mathematics textbook. Fifth, in the New Zealand textbook they had used teaching methods inducing curiosity of learner such as finding message and puzzle problem than solving given problem simply.

Association of Genetic Polymorphism of IL-2 Receptor Subunit and Tuberculosis Case

  • Lee, Sang-In;Jin, Hyun-Seok;Park, Sangjung
    • Biomedical Science Letters
    • /
    • v.24 no.2
    • /
    • pp.94-101
    • /
    • 2018
  • Tuberculosis (TB) is infectious disease caused by Mycobacterium tuberculosis (MTB) infection. It is known that not only the property of microorganism but also the genetic susceptibility of infected patients is controlled. Interleukin 2 (IL-2) is a cytokine belonging to type 1 T helper (Th1) activity. In addition, IL-2, when infected with MTB, binds IL-2 receptor and promotes T cell replication and is involved in granuloma formation. The aim of this study was to investigate the genetic polymorphisms of the IL-2 receptor gene in tuberculosis patients and normal individuals. We analyzed 22 SNPs in three genes using the genotype data of 443 tuberculosis cases and 3,228 healthy controls from the Korea Association Resource for their correlation with tuberculosis case. IL2RA, IL2RB, and IL2RG genes were genotyped of 16, 4, and 2 SNPs, respectively. Among three genes, only IL2RA gene polymorphisms showed statistically significant association with tuberculosis case. 6 SNPs with high significance were identified in the IL2RA gene. In addition, the linkage disequilibrium (LD) structure of IL2RA gene was confirmed. SNP imputation of IL2RA gene was performed, it was confirmed that more SNPs were significant between case and control. If we look at the results of IL2RA gene analysis above, we can see that genetic polymorphism in the gene expressing $IL-2R{\alpha}$ will regulate the expression level of $IL-2R{\alpha}$, and the change in the immune system involved in $IL-2R{\alpha}$. In this study, genetic polymorphism that may affect host immunity suggests that susceptibility to tuberculosis may be controlled.

Export of Human Proinsulin in E. coli : High Export of Proinsulin Fusion Protein but not of Proinsulin Itself (대장균에서 인체 프로인슐린의 분비 발현 : 프로인슐린 융합체의 고분비 발현과 프로인슐린의 저분비 발현)

  • Yup Kang
    • KSBB Journal
    • /
    • v.11 no.2
    • /
    • pp.165-172
    • /
    • 1996
  • To obtain a correctly folded human proinsulin, export of proinsulin using Staphylococcal protein A signal sequence-mediated secretion pathway has been attempted in E.coli. A secretion operon for proinsulin was constructed by consecutively connecting T7 promoter, SPA ribosome binding site, SPA signal sequence gene, and human proinsulin gene. Little immunoreactive proinsulin was detected in the periplasmic space and. culture medium, and not even in cytoplasmic space. The qualitative analysis of transcribed proinsulin mRNA and the in vitro transcription/translation experiment suggests that the negligible level of proinsulin export appears to be due to intracellular degradation of proinsulin, rather than due to the blockage during translocation. However, expression of proinsulin fusion protein such as MBP-proinsulin could dramatically increase export of proinsulin in E.coli.

  • PDF

Tubular Jacquard Knit Wear Design through the Application of Keith Haring's Works (키스 해링[Keith Haring]의 작품을 응용한 니트웨어 디자인 - 튜블러 자카드 조직을 활용하여 -)

  • Lee, Ha-Jung;Lee, Youn-Hee
    • Journal of the Korean Society of Costume
    • /
    • v.61 no.1
    • /
    • pp.69-83
    • /
    • 2011
  • This study applied cartoon-like simple and implicit symbol of Keith Haring works to tubular jacquard knit, designed woman casual knit wear with fun, explicit and activities using fluorescent yarns with explicit features. This study aimed to propose the directions for new market pioneering that meets the emotional requirements of modern people by raising the level of utilization and expression area of knit fashion, and to present the constructive directions for high value-added knit wear by designing highly effective knit wear. Methods of this study examined the lifetime and artistic background of Keith Haring and collected his works through searching of internet, scholastic publications or thesis. Formative types and colors of works collected were analyzed and classified according to characteristic standards of Haring and characteristic formativeness and color of works were extracted. The result of this study is as shown below: First, the feature of his works can be summarized into motif that has symbolic and implicit cartoon-like lines including narratives as well as enormous number of works. Such feature is very suitable for variation and recombination in realization of design and has unlimited potentials for development. Second, image colors can be extracted such as black and white, original color and fluorescent paints, which reflects the intention of artist who put focus on communication with the public. Such color feature is very suitable for design motif that has explicitness of reflecting the intention of artist through use of special fluorescent yarn. Third, I made it as reversible wear that has effects of increasing or decreasing the explicitness according to color area ratio using the feature of textures allowing use of both sides as well as form stability of tubular jacquard. Fourth, by adjusting the thickness of fabric with controlling of ply in fabric using poly yarn and wool union yarn, I could obtain good results of study in terms of technique that can express various materials with embossed dynamic effects of unevenness on flat surface.

Proteomic Characterization of the 'Agakong', a Small-seeded Recombinant Inbred Line Derived from 'Eunhakong' (Glycine max) $\times$ 'KLG10084' (Glycine soja)

  • Choi, Ung-Kyu;Ryu, Hyun-Su;Kim, Hyun-Tae;Yun, Sun-Mi;Lee, Su-Jin;Choi, Jae-Dek;Hwang, Young-Hyun;Choi, Soo-Young;Kwon, Oh-Shin
    • Food Science and Biotechnology
    • /
    • v.17 no.5
    • /
    • pp.912-918
    • /
    • 2008
  • This study was conducted to identify the differences in proteomic characteristics of 'Agakong', recombinant inbred line, and its parental genotypes 'Eunhakong' (Glycine max) and 'KLG10084' (G. soja). The isoflavone content of 'Agakong' was 3 times higher than that of its parental lines. A combined high-throughput proteomic approach was employed to determine the expression profile and identity of proteins using 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. The overall distribution patterns of proteins are quite similar, but lots of protein spot intensities varied among the genotypes. A total of 41 proteins, representing significant difference in the quantities of protein among the lines, were successfully identified. Among them, more than 50% of the proteins identified were subunits of glycinin and $\beta$-conglycinin, 2 major storage proteins. This study showed that the proteomic analysis could help to define specific changes in protein level and composition, which can occur in the generation of new soybean varieties.

Proteomic Characteristics of Calcium Enriched King Oyster Mushroom (Pleurotus eryngii) (칼슘함량이 강화된 새송이 버섯의 프로테옴 분석)

  • Bae, Hee-Sun;Kim, Dae-Hyun;Choi, Ung-Kyu
    • Korean Journal of Food Science and Technology
    • /
    • v.43 no.1
    • /
    • pp.12-16
    • /
    • 2011
  • This study was conducted to identify the differences in proteomic characteristics between Ca-enriched king oyster mushrooms and general king oyster mushrooms. A combined high-throughput proteomic approach was employed to determine the expression profiles and identity of proteins using 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. The overall distribution patterns of the proteins were quite similar, but many of the protein spot intensities varied. A total of 10 proteins, representing a significant difference in the quantities of protein betweenthe two types of mushrooms, were successfully identified. Among these proteins, eight kinds were increased in the Ca-enriched king oyster mushrooms and two kinds were decreased. This study showed that proteomic analysis can help define specific changes in protein level and composition, which can occur in mushrooms where Ca content may or may not be enriched.

OsATG10b, an Autophagosome Component, Is Needed for Cell Survival against Oxidative Stresses in Rice

  • Shin, Jun-Hye;Yoshimoto, Kohki;Ohsumi, Yoshinori;Jeon, Jong-seong;An, Gynheung
    • Molecules and Cells
    • /
    • v.27 no.1
    • /
    • pp.67-74
    • /
    • 2009
  • Autophagy degrades toxic materials and old organelles, and recycles nutrients in eukaryotic cells. Whereas the studies on autophagy have been reported in other eukaryotic cells, its functioning in plants has not been well elucidated. We analyzed the roles of OsATG10 genes, which are autophagy-related. Two rice ATG10 genes - OsATG10a and OsATG10b - share significant sequence homology (about 75%), and were ubiquitously expressed in all organs examined here. GUS assay indicated that OsATG10b was highly expressed in the mesophyll cells and vascular tissue of younger leaves, but its level of expression decreased in older leaves. We identified T-DNA insertional mutants in that gene. Those osatg10b mutants were sensitive to treatments with high salt and methyl viologen (MV). Monodansylcadaverine-staining experiments showed that the number of autophagosomes was significantly decreased in the mutants compared with the WT. Furthermore, the amount of oxidized proteins increased in MV-treated mutant seedlings. These results demonstrate that OsATG10b plays an important role in the survival of rice cells against oxidative stresses.

Immunization with a Genetically Engineered Uropathogenic Escherichia coli Adhesin-Escherichia coli Enterotoxin Subunit A2B Chimeric Protein

  • Lee, Yong-Hwa;Kim, Byung-O;Pyo, Suhk-Neung
    • Biomolecules & Therapeutics
    • /
    • v.13 no.2
    • /
    • pp.101-106
    • /
    • 2005
  • The generation of secretory IgA antibodies (Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the heat-labile Escherichia coli enterotoxin A2B (ltxa2b) gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ltxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and westem blotting using antibodies to the maltose binding protein (MBP) or the heat labile E. coli subunit B (LTXB), plus the N-terminal amino acid sequence was analyzedd. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/LTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of LTXB. thisstudy also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA (sIgA) and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked LTXA2B acts as a useful mucosal adjuvant, and that adhesin/LTXA2A chimeric protein might be a potential antigen for oral immunization against UPEC.

One-step spectral clustering of weighted variables on single-cell RNA-sequencing data (단세포 RNA 시퀀싱 데이터를 위한 가중변수 스펙트럼 군집화 기법)

  • Park, Min Young;Park, Seyoung
    • The Korean Journal of Applied Statistics
    • /
    • v.33 no.4
    • /
    • pp.511-526
    • /
    • 2020
  • Single-cell RNA-sequencing (scRNA-seq) data consists of each cell's RNA expression extracted from large populations of cells. One main purpose of using scRNA-seq data is to identify inter-cellular heterogeneity. However, scRNA-seq data pose statistical challenges when applying traditional clustering methods because they have many missing values and high level of noise due to technical and sampling issues. In this paper, motivated by analyzing scRNA-seq data, we propose a novel spectral-based clustering method by imposing different weights on genes when computing a similarity between cells. Assigning weights on genes and clustering cells are performed simultaneously in the proposed clustering framework. We solve the proposed non-convex optimization using an iterative algorithm. Both real data application and simulation study suggest that the proposed clustering method better identifies underlying clusters compared with existing clustering methods.