• Title/Summary/Keyword: ENCODE

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Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.415-422
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    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Histone Methylation Regulates Retinoic Acid-induced Hoxc Gene Expression in F9 EC Cells (F9 EC 세포에서 레티노산에 의해 유도되는 Hoxc 유전자의 발현에 히스톤 메틸화가 미치는 영향)

  • Min, Hyehyun;Kim, Myoung Hee
    • Journal of Life Science
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    • v.25 no.6
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    • pp.703-708
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    • 2015
  • Hox genes encode a highly conserved family of homeodomain-containing transcription factors controlling vertebrate pattern formation along the anteroposterior body axis during embryogenesis. Retinoic acid (RA) is a key morphogen in embryogenesis and a critical regulator of both adult and embryonic cellular activity. Specifically, RA regulates Hox gene expression in mouse- or human-derived embryonic carcinoma (EC) cells. Histone modification has been reported to play a pivotal role in the process of RA-induced gene expression and cell differentiation. As histone modification is thought to play an essential role in RA-induced Hox gene expression, we examined RA-induced initiation of collinear expression of Hox genes and the corresponding histone modifications in F9 murine embryonic teratocarcinoma (EC) cells. Hox expression patterns and histone modifications were analyzed by semiquantitative RT-PCR, RNA-sequencing, and chromatin immuno-precipitation (ChIP)-PCR analyses. The Hoxc4 gene (D0) was initiated earlier than the Hoxc5 to –c10 genes (D3) upon RA treatment (day 0 [D0], day 1 [D1], and day 3 [D3]). The Hox nonexpressing D0 sample had a strong repressive marker, H3K27me3, than the D1 and D3 samples. In the D1 and D3 samples, reduced enrichment of the H3K27me3 marker was observed in the whole cluster. The active H3K4me3 marker was closely associated with the collinear expression of Hoxc genes. Thus, the Hoxc4 gene (D1) and all Hoxc genes (D3) expressed H3K4me3 upon transcription activation. In conclusion, these data indicated that removing H3K27me3 and acquiring H3K4me3 regulated RA-induced Hoxc gene collinearity in F9 cells.

Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Recognizing the Direction of Action using Generalized 4D Features (일반화된 4차원 특징을 이용한 행동 방향 인식)

  • Kim, Sun-Jung;Kim, Soo-Wan;Choi, Jin-Young
    • Journal of the Korean Institute of Intelligent Systems
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    • v.24 no.5
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    • pp.518-528
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    • 2014
  • In this paper, we propose a method to recognize the action direction of human by developing 4D space-time (4D-ST, [x,y,z,t]) features. For this, we propose 4D space-time interest points (4D-STIPs, [x,y,z,t]) which are extracted using 3D space (3D-S, [x,y,z]) volumes reconstructed from images of a finite number of different views. Since the proposed features are constructed using volumetric information, the features for arbitrary 2D space (2D-S, [x,y]) viewpoint can be generated by projecting the 3D-S volumes and 4D-STIPs on corresponding image planes in training step. We can recognize the directions of actors in the test video since our training sets, which are projections of 3D-S volumes and 4D-STIPs to various image planes, contain the direction information. The process for recognizing action direction is divided into two steps, firstly we recognize the class of actions and then recognize the action direction using direction information. For the action and direction of action recognition, with the projected 3D-S volumes and 4D-STIPs we construct motion history images (MHIs) and non-motion history images (NMHIs) which encode the moving and non-moving parts of an action respectively. For the action recognition, features are trained by support vector data description (SVDD) according to the action class and recognized by support vector domain density description (SVDDD). For the action direction recognition after recognizing actions, each actions are trained using SVDD according to the direction class and then recognized by SVDDD. In experiments, we train the models using 3D-S volumes from INRIA Xmas Motion Acquisition Sequences (IXMAS) dataset and recognize action direction by constructing a new SNU dataset made for evaluating the action direction recognition.

Digital Hologram Compression Technique By Hybrid Video Coding (하이브리드 비디오 코팅에 의한 디지털 홀로그램 압축기술)

  • Seo, Young-Ho;Choi, Hyun-Jun;Kang, Hoon-Jong;Lee, Seung-Hyun;Kim, Dong-Wook
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.42 no.5 s.305
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    • pp.29-40
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    • 2005
  • According as base of digital hologram has been magnified, discussion of compression technology is expected as a international standard which defines the compression technique of 3D image and video has been progressed in form of 3DAV which is a part of MPEG. As we can identify in case of 3DAV, the coding technique has high possibility to be formed into the hybrid type which is a merged, refined, or mixid with the various previous technique. Therefore, we wish to present the relationship between various image/video coding techniques and digital hologram In this paper, we propose an efficient coding method of digital hologram using standard compression tools for video and image. At first, we convert fringe patterns into video data using a principle of CGH(Computer Generated Hologram), and then encode it. In this research, we propose a compression algorithm is made up of various method such as pre-processing for transform, local segmentation with global information of object image, frequency transform for coding, scanning to make fringe to video stream, classification of coefficients, and hybrid video coding. Finally the proposed hybrid compression algorithm is all of these methods. The tool for still image coding is JPEG2000, and the toots for video coding include various international compression algorithm such as MPEG-2, MPEG-4, and H.264 and various lossless compression algorithm. The proposed algorithm illustrated that it have better properties for reconstruction than the previous researches on far greater compression rate above from four times to eight times as much. Therefore we expect that the proposed technique for digital hologram coding is to be a good preceding research.

Development of Genetic Selection Marker via Examination of Genome in Bacillus velezensis K10 (Bacillus velezensis K10 유전체 분석을 통한 균주 선발 마커 개발)

  • Sam Woong Kim;Young Jin Kim;Tae Wook Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.33 no.11
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    • pp.897-904
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    • 2023
  • This study was done to develope genetic markers with the unique characteristics of genes according to the genomic information of Bacillus velezensis K10. B. velezensis K10 maintained a total of 4,159,835 bps, which was found to encode 5,136 open reading frames (orfs). B. velezensis K10 was found to have much more gene migration due to external factors overall compared to standard strain B. velezensis JS25R. In order to discover genetic selection markers, orfs on the genome to be easily induced to gene mutation were surveyed such as recombinase, integrase, transposase, and phage-related genes. As a result of the investigation, 9 candidate markers were isolated with high possibility as genetic selection markers. Although a part in the various origin's areas showed specificities in comparison with homology, the selected markers were all existed in phage-related areas because they were relatively lower homologies in phage-related genes. PCR analysis was done on B. licheniformis K12, B. velezensis K10, B. subtilis, and B. cereus to establish them as inter-species candidate selection markers. As a result, it was confirmed that B. velezensis K10-specific PCR products were formed in a total of 6 primer sets such as BV3 and BV5 to 9. On the other hand, analysis at the subspecies level observed the formation of B. velezensis K10-specific PCR products in 4 primer sets such as BV3, 5, 8, and 9. Among them, since BV5 and BV8 were detected by very specific results, we suggest that BV5 and 8 can be used as B. velezensis K10 gene selection markers at the species and sub-species level.