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The multifunctional RNA-binding protein hnRNPK is critical for the proliferation and differentiation of myoblasts

  • Xu, Yongjie;Li, Rui;Zhang, Kaili;Wu, Wei;Wang, Suying;Zhang, Pengpeng;Xu, Haixia
    • BMB Reports
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    • v.51 no.7
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    • pp.350-355
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    • 2018
  • HnRNPK is a multifunctional protein that participates in chromatin remodeling, transcription, RNA splicing, mRNA stability and translation. Here, we uncovered the function of hnRNPK in regulating the proliferation and differentiation of myoblasts. hnRNPK was mutated in the C2C12 myoblast cell line using the CRISPR/Cas9 system. A decreased proliferation rate was observed in hnRNPK-mutated cells, suggesting an impaired proliferation phenotype. Furthermore, increased G2/M phase, decreased S phase and increased sub-G1 phase cells were detected in the hnRNPK-mutated cell lines. The expression analysis of key cell cycle regulators indicated mRNA of Cyclin A2 was significantly increased in the mutant myoblasts compared to the control cells, while Cyclin B1, Cdc25b and Cdc25c were decreased sharply. In addition to the myoblast proliferation defect, the mutant cells exhibited defect in myotube formation. The myotube formation marker, myosin heavy chain (MHC), was decreased sharply in hnRNPK-mutated cells compared to control myoblasts during differentiation. The deficiency in hnRNPK also resulted in the repression of Myog expression, a key myogenic regulator during differentiation. Together, our data demonstrate that hnRNPK is required for myoblast proliferation and differentiation and may be an essential regulator of myoblast function.

Automatic Target Recognition by selecting similarity-transform-invariant local and global features (유사변환에 불변인 국부적 특징과 광역적 특징 선택에 의한 자동 표적인식)

  • Sun, Sun-Gu;Park, Hyun-Wook
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.39 no.4
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    • pp.370-380
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    • 2002
  • This paper proposes an ATR (Automatic Target Recognition) algorithm for identifying non-occluded and occluded military vehicles in natural FLIR (Forward Looking InfraRed) images. After segmenting a target, a radial function is defined from the target boundary to extract global shape features. Also, to extract local shape features of upper region of a target, a distance function is defined from boundary points and a line between two extreme points. From two functions and target contour, four global and four local shape features are proposed. They are much more invariant to translation, rotation and scale transform than traditional feature sets. In the experiments, we show that the proposed feature set is superior to the traditional feature sets with respect to the similarity-transform invariance and recognition performance.

Concatemer-Associated Transgene Expression Patterns in Transgenic Marine Medaka Oryzias dancena Strains

  • Cho, Young Sun;Kim, Dong Soo;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • v.18 no.1
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    • pp.73-80
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    • 2015
  • To examine the interrelationship between transgenic insertion patterns and transgene expression profiles in established transgenic fish lines, four stable transgenic marine medaka Oryzias dancena germlines harboring ${\beta}$-actin regulator-driven RFP reporter constructs were selected. The established transgenic strains were characterized with regard to their transgenic genotypes (insertion pattern, concatemer formation, and transgene copy number based on genomic Southern blot hybridization and qPCR assay) and expression characteristics at the mRNA (qRT-PCR), protein (western blot), and phenotypic (fluorescent appearance) levels. From comparative examinations, it was found that transgenic expression at both the transcription and translation levels could be significantly downregulated in transgenic strains, potentially through methylation-mediated transgene silencing that was particularly associated with the formation of a long tail-to-head tandem concatemer in the chromosomal integration site(s). When this occurred, an inverse relationship between the transgene copy number and fluorescence intensity was observed in the resultant transgenic fish. However, with the other transgenic genotype, transgenic individuals with an identical Southern blot hybridization pattern, containing a tandem concatemer(s), had very different expression levels (highly robust vs. low expression strengths), which was possibly related to the differential epigenetic modifications and/or degrees of methylation. The concatemer-dependent downregulation of transgene activity could be induced in transgenic fish, but the overall pattern was strain-specific. Our data suggest that neither a low (or single) transgene copy number nor tandem transgene concatemerization is indicative of strong or silenced transgene expression in transgenic fish carrying a ubiquitous transgene. Hence, a sufficient number of transgenic lineages, with different genotypes, should be considered to ensure the establishment of the best-performance transgenic line(s) for practical applications.

Zebrafish Dnd protein binds to 3'UTR of geminin mRNA and regulates its expression

  • Chen, Shu;Zeng, Mei;Sun, Huaqin;Deng, Wenqian;Lu, Yilu;Tao, Dachang;Liu, Yunqiang;Zhang, Sizhong;Ma, Yongxin
    • BMB Reports
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    • v.43 no.6
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    • pp.438-444
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    • 2010
  • Dnd (dead end) gene encodes an RNA binding protein and is specifically expressed in primordial germ cells (PGCs) as a vertebrate-specific component of the germ plasma throughout embryogenesis. By utilizing a technique of specific nucleic acids associated with proteins (SNAAP), 13 potential target mRNAs of zebrafish Dnd (ZDnd) protein were identified from 8-cell embryo, and 8 target mRNAs have been confirmed using an RT-PCR analysis. Of the target mRNAs, the present study is focused on the regulation of geminin, which is an inhibitor of DNA replication. Using electrophoretic mobility shift assay (EMSA), we demonstrated that ZDND protein bound the 67-nucleotide region from 864 to 931 in the 3'UTR of geminin mRNA, a sequence containing 60.29% of uridine. Results from a dual-luciferase assay in HEK293 cells showed that ZDND increases the translation of geminin. Taken together, the identification of target mRNA for ZDnd will be helpful to further explore the biological function of Dnd in zebrafish germ-line development as well as in cancer cells.

Online Handwritten Digit Recognition by Smith-Waterman Alignment (Smith-Waterman 정렬 알고리즘을 이용한 온라인 필기체 숫자인식)

  • Mun, Won-Ho;Choi, Yeon-Seok;Lee, Sang-Geol;Cha, Eui-Young
    • Journal of the Korea Society of Computer and Information
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    • v.16 no.9
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    • pp.27-33
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    • 2011
  • In this paper, we propose an efficient on-line handwritten digit recognition base on Convex-Concave curves feature which is extracted by a chain code sequence using Smith-Waterman alignment algorithm. The time sequential signal from mouse movement on the writing pad is described as a sequence of consecutive points on the x-y plane. So, we can create data-set which are successive and time-sequential pixel position data by preprocessing. Data preprocessed is used for Convex-Concave curves feature extraction. This feature is scale-, translation-, and rotation-invariant. The extracted specific feature is fed to a Smith-Waterman alignment algorithm, which in turn classifies it as one of the nine digits. In comparison with backpropagation neural network, Smith-Waterman alignment has the more outstanding performance.

Regulation of HMG-CoA Reductase mRNA Stability by 25-hydroxycholesterol

  • Park, Jae-Won;Oh, Seung-Min
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.184-188
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    • 2000
  • HMG-CoA reductase is th rate-limiting enzyme of cholesterol biosynthesis. As intracellular levels of cholesterol should be regulated elaborately in response to external stimuli an internal needs, the expression of the HMG-CoA reductase gene is regulated intricately at several different levels from transcription to post-translational modification. In this study, we investigated the regulatory mechanism of HMG-CoA reductase gene expression at the post-transcriptional/pre-translational levels in a baby hamster kidney cell line, C100. when 25-hydroxycholesterol was added to cells cultured in medium containing 5% delipidized fetal bovine serum and 25$\mu$M lovastatin, the levels of HMG-CoA reductase mRNA decreased rapidly, which seemed to be due to the increased degradation of reductase mRNA. These suppressive effects of 25-hydroxycholesterol on MG-CoA reductase mRNA levels were blocked by a translation inhibitor, cycloheximide. Similarly, actinomycin D and 5,6-dichloro-1-$\beta$-D-ribofuranosylbenzimidazole, transcription inhibitors, blocked the 25-hydroxycholesterol-mediated degradation of HMG-CoA reductase mRNA. These results indicate that new protein/RNA synthesis is required for the degradation of HMG-CoA reductase mRNA. In addition, data from the transfection experiments shows that cis-acting determinants, regulating the stability of reductase mRNA, were scattered in the sequence corresponding to 1766-4313 based on the sequence of Syrian hamster HMG-CoA reductase cDNA. Our data suggests that sterol-mediated destabilization of reductase mRNA might be one of the important regulatory mechanism of HMG-CoA reductase gene expression.

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Realistic 3D model generation of a real product based on 2D-3D registration (2D-3D 정합기반 실제 제품의 사실적 3D 모델 생성)

  • Kim, Gang Yeon;Son, Seong Min
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.11
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    • pp.5385-5391
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    • 2013
  • As on-line purchases is activated, customers' demand increases for the realistic and accurate digital information of a product design. In this paper, we propose a practical method that can generate a realistic 3D model of a real product using a 3D geometry obtained by a 3D scanner and its photographic images. In order to register images to the 3D geometry, the camera focal length, the CCD scanning aspect ratio and the transformation matrix between the camera coordinate and the 3D object coordinate must be determined. To perform this 2D-3D registration with consideration of computational complexity, a three-step method is applied, which consists of camera calibration, determination of a temporary optimum translation vector (TOTV) and nonlinear optimization for three rotational angles. A case study for a metallic coated industrial part, of which the colour appearance is hardly obtained by a 3D colour scanner has performed to demonstrate the effectiveness of the proposed method.

Active Shape Model-based Objectionable Image Detection (활동적 형태 모델을 이용한 유해영상 탐지)

  • Jang, Seok-Woo;Joo, Seong-Il;Kim, Gye-Young
    • Journal of Internet Computing and Services
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    • v.10 no.5
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    • pp.183-194
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    • 2009
  • In this paper, we propose a new method for detecting objectionable images with an active shape model. Our method first learns the shape of breast lines through principle component analysis and alignment as well as the distribution of intensity values of corresponding landmarks, and then extracts breast lines with the learned shape and intensity distribution. To accurately select the initial position of active shape model, we obtain parameters on scale, rotation, and translation. After positioning the initial location of active shape model using scale and rotation information, iterative searches are performed. We can identify adult images by calculating the average of the distance between each landmark and a candidate breast line. The experiment results show that the proposed method can detect adult images effectively by comparing various results.

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A Study on the Radiation Characteristics of Microstrip Array Antennas on the Nonplanar Surface (곡면에서의 마이크로스트립 어레이 안테나의 복사 특성에 관한 연구)

  • 구연건;이정수;고광태
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.14 no.2
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    • pp.121-136
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    • 1989
  • In this paper, an attempt has been made to analyze the theoretically and verify experimentally the effect of curvature on the radiation characteristics of microstrip array antennas mounted conformally on the concave surface and the convex surface of the cylindrical body. The analysis of single element microstrip antenna is made by using the analysis method of Transmission Line Model. The theory of array antennas is established by application of the method of transformed coordinates, in which the translation and the ratation about each single element arrayed two-demensionally on the nonplanar surface are under consideration, and it is investigated by computation of the synthetic electric field strength in the far zone. In addition, various radiation characteristics, such as return loss, resonant frequency, radiation pattern, half-power, beamwidth, gain, are measrued and compared with the theroetical values according to the variation of curvature, by designing and building 4-element array microstrip antenna operating at 10 GHz, and microstrip feed lines. As predicted in theroy, it is verified that radiation pattern of antennas mounted on the concave and the convex surfaces alike broadens as the radius of curvature decreases. And for the curved surfaces, aggrement between computed values of the total synthetic radiation power pattern by the method of transformed coordinates and measured valuse is good. Besides, it is found that resonant frequency, input impedance and gain are hardly affected by the radius of curvature.

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Expression Profiling of Lipopolysaccharide Target Genes in RAW264.7 Cells by Oligonucleotide Microarray Analyses

  • Huang, Hao;Park, Cheol-Kyu;Ryu, Ji-Yoon;Chang, Eun-Ju;Lee, Young-Kyun;Kang, Sam-Sik;Kim, Hong-Hee
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.890-897
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    • 2006
  • In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.