• Title/Summary/Keyword: Genome Analysis

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Molecular Phylogeny of Korean-type Coliphages and American-type Coliphages Determined by a RAPD Analysis (RAPD 분석법에 의한 한국형 대장균파아지와 미국형 대장균파아지의 분자적 계통분류)

  • 권오식
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.131-139
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    • 2000
  • RAPD-PCR was applied to identify the phylogenetic relationship between isolated Korean-type coliphages ($\phi$C1, $\phi$C2, $\phi$C3 and $\phi$C4) and well-known American coliphages ($\phi$T2, $\phi$T4, $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$). Subsequently, a computer analysis was carried out with the results of RAPD-PCR. As a result, 9 individuals were divided into five groups. The Korean-type coliphages formed a single cluster which showed very high genetic similarity but the American-type coliphages revealed very low genetic similarity among them. In particular, the $\phi$T2와 $\phi$T4 (T$_{even}$ phages) made one sub-cluster among American coliphages, and they were very distant from $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$. However, ${\phi}{\lambda}$ made a cluster with the Korean-type coliphages that we isolated. The genome size of Korean-type coliphages was ranged from 25,000 bp to 35,000 bp. Among them, the genome of $\phi$C2 was the smallest and that of $\phi$C1 was the biggest, while others were in the middle of the size.

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Transcription Factor for Gene Function Analysis in Maize (옥수수 유전자 기능 분석을 위한 전사인자의 이해)

  • Moon, Jun-Cheol;Kim, Jae Yoon;Baek, Seong-Bum;Kwon, Young-Up;Song, Kitae;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.263-281
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    • 2014
  • Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3/VP1, AP2/EREBP, ARF, ARID, AS2, AUX/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2-YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.

Cloning and Sequence Analysis of the Full-length cDNA of Coxsackievirus B3 Isolated in Korea (한국에서 분리된 콕사키 바이러스 B3 cDNA의 클로닝 및 전체 염기서열 분석)

  • Chung, Yoon-Seok;Kim, Ki-Soon;Park, Jeong-Koo;Lee, Yoon-Sung;Shin, Soo-Youn;Cheon, Doo-Seong;Jee, Young-Mee;Kim, Moon-Bo;Na, Byoung-Kuk;Yoon, Jae-Deuk;Lee, Kwang-Ho;Song, Chul-Yong
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.71-81
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    • 2000
  • We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.

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Sequence Analysis of Small Round Structured Viruses (SRSV) Isolated from a Diarrheal Patient in Wonju (원주지역 설사 환자에서 분리한 Small Round Structured Viruses (SRSV) 염기서열 분석)

  • Jee, Young-Mee;Kim, Ki-Soon;Cheon, Doo-Sung;Park, Jeong-Koo;Kang, Young-Hwa;Chung, Yoon-Suck;Go, Un-Yeong;Shin, Young-Hack;Yoon, Jae-Deuk
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.247-259
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    • 1999
  • Small round structured viruses (SRSV) are the major ethological agents which can cause outbreaks of non-bacterial gastroenteritis or food poisoning both in children and adults. The classification of family Caliciviridae to which SRSV belong, is based on the genome encoding three open reading frames. The rotavirus is another major pathogen which causes diarrhea in young children. We examined stool specimens obtained from diarrheal patients in Wonju from which bacterial pathogens were not found. To detect causative viruses from stool specimens of patients, reverse transcription (RT)-polymerase chain reaction (PCR) or nested PCR using rotavirus or SRSV specific primers was performed. In this study, RT-nested PCR procedure which can amplify a 330 bp fragment derived from RNA dependent RNA polymerase (RDRP) region within ORF1 was applied for the detection of SRSV. For the detection of rotaviruses, a 877 bp fragment from the VP4 region of rotavirus genome was amplified. As a result, rotavirus was not detected while SRSV sequences were detected from one out of five specimens. The nucleotide and amino acid sequences of the Wonju isolate were compared with other 6 Korean isolates which have been isolated and sequenced in our laboratory. Sequence analysis revealed that the Wonju isolate was rather distinct from other Korean isolates: the Wonju isolate was closer to genogroup I of SRSV while other 6 Korean isolates belonged to genogroup II.

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Isolation of Circadian-associated Genes in Brassica rapa by Comparative Genomics with Arabidopsis thaliana

  • Kim, Jin A;Yang, Tae-Jin;Kim, Jung Sun;Park, Jee Young;Kwon, Soo-Jin;Lim, Myung-Ho;Jin, Mina;Lee, Sang Choon;Lee, Soo In;Choi, Beom-Soon;Um, Sang-Hee;Kim, Ho-Il;Chun, Changhoo;Park, Beom-Seok
    • Molecules and Cells
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    • v.23 no.2
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    • pp.145-153
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    • 2007
  • Elucidation of the roles of circadian associated factors requires a better understanding of the molecular mechanisms of circadian rhythms, control of flowering time through photoperiodic pathways, and photosensory signal transduction. In Arabidopsis, the APRR1 quintet, APRRs 1, 3, 5, 7, and 9, are known as central oscillator genes. Other plants may share the molecular mechanism underlying the circadian rhythm. To identify and characterize these circadian response genes in Brassica crops whose genome was triplicated after divergence from Arabidopsis, we identified B. rapa BAC clones containing these genes by BLAST analysis of B. rapa BAC end sequences against the five corresponding Arabidopsis regions. Subsequent fingerprinting, Southern hybridization, and PCR allowed identification of five BAC clones, one for each of the five circadian-related genes. By draft shotgun sequencing of the BAC clones, we identified the complete gene sequences and cloned the five expressed B. rapa circadian-associated gene members, BrPRRs 1, 3, 5, 7, and 9. Phylogenetic analysis revealed that each BrPRR was orthologous to the corresponding APRR at the sequence level. Northern hybridization revealed that the five genes were transcribed at distinct points in the 24 hour period, and Southern hybridization revealed that they are present in 2, 1, 2, 2, and 1 copies, respectively in the B. rapa genome, which was triplicated and then diploidized during the last 15 million years.

A Molecular Study of Rice Black-Streaked Dwarf Virus (벼 흑조위축병 바이러스의 분자생물학적 연구)

  • Park, Jong-Sug;Bae, Shin-Chyul;Kim, Young-Min;Paik, Young-Ki;Kim, Ju-Kon;Hwang, Young-Soo
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.148-153
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    • 1994
  • Rice black-streaked dwarf virus (RBSDV), a member of the plant reoviridae fijivirus group, causes a serious damage for rice production in Korea. To characterize the RBSDV genome, virus particles were produced by feeding of planthopper (Laodelphax striatellus F.) carring RBSDV to maize plants for 2 days. In $30{\sim}40$ days after feeding, the viral particles were purified from the infected maize roots by using $10{\sim}40%$ sucrose gradient centrifugation. After treatment of 10% SDS to remove the viral coat proteins, ten viral double-stranded RNAs were resolved in agrose gel electrophoresis. Total dsRNA was then used to synthesize cDNA by reverse transcriptase and a cDNA library was constructed in the ${\lambda}gt11$ vector. The phages that contain RBSDV cDNA fragments were selected by hybridizing with the random-primed probe prepared from RBSDV dsRNAs. After subcloning of several cDNA fragments into the pUC19 plasmid vector, one clone (pRV3) was chosen for sequencing. The pRV3 clone was shown to be located on the RBSDV genome fragment No.3 by RNA gel-blot analysis. Sequence analysis of the clone revealed that the pRV3 contains two partial open reading frames.

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Genome-based Gene Expression Analysis of EGCG-mediated Cell Transformation Suppression Effect in Mouse Cell line Balb/c 3T3 A31-1-1 (마우스세포주 Balb/c 3T3 A31-1-1에서 Epigallocatechin gallate(EGCG)의 세포암화 억제효과에 대한 유전자발현 해석)

  • Jung, Ki-Kyung;Suh, Soo-Kyung;Kim, Tae-Gyun;Park, Moon-Suk;Lee, Woo-Sun;Park, Sue-Nie;Kim, Seung-Hee;Jung, Hai-Kwan
    • Environmental Mutagens and Carcinogens
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    • v.26 no.4
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    • pp.125-132
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    • 2006
  • Previous studies showed that epigallocatechin gallate(EGCG) have substantial effects of suppressing the N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)-initiated cell transformation process on the bases of foci formation frequency and loss of anchorage dependency. In this study we tried to clarify the molecular mechanism of suppressing the cell transformation process. Mouse cell line balb/c 3T3 A31-1-1 was exposed 2 days to MNNG followed by 15 days 12-O-tetradecanoylphorbol-13-acetate(TPA) treatment for our transformation process. EGCG was added after the time point of 24 hours exposure to TPA and incubated for 19 days. 2029 genes were selected in our transformation process that showed fold change value of 1.5 or more in the microarray gene expression analysis covering the mouse full genome. These genes were found to be involved mainly in the cell cycle pathway, focal adhesion, adherens junction, TGE-$\beta$ signaling, apoptosis, lysine degradation, insulin signaling, ECM-receptor interaction. Among the genes, we focused on the 631 genes(FC>0.5) reciprocally affected by EGCG treatment. Our study suggest that EGCG down-regulate the gene expressions of up stream signaling factors such as nemo like kinase with MAPK activity and PI3-Kinase, Ras GTPase and down stream factors such as cyclin D1, D2, H, T2, cdk6.

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Direct Pass-Through based GPU Virtualization for Biologic Applications (바이오 응용을 위한 직접 통로 기반의 GPU 가상화)

  • Choi, Dong Hoon;Jo, Heeseung;Lee, Myungho
    • KIPS Transactions on Software and Data Engineering
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    • v.2 no.2
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    • pp.113-118
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    • 2013
  • The current GPU virtualization techniques incur large overheads when executing application programs mainly due to the fine-grain time-sharing scheduling of the GPU among multiple Virtual Machines (VMs). Besides, the current techniques lack of portability, because they include the APIs for the GPU computations in the VM monitor. In this paper, we propose a low overhead and high performance GPU virtualization approach on a heterogeneous HPC system based on the open-source Xen. Our proposed techniques are tailored to the bio applications. In our virtualization framework, we allow a VM to solely occupy a GPU once the VM is assigned a GPU instead of relying on the time-sharing the GPU. This improves the performance of the applications and the utilization of the GPUs. Our techniques also allow a direct pass-through to the GPU by using the IOMMU virtualization features embedded in the hardware for the high portability. Experimental studies using microbiology genome analysis applications show that our proposed techniques based on the direct pass-through significantly reduce the overheads compared with the previous Domain0 based approaches. Furthermore, our approach closely matches the performance for the applications to the bare machine or rather improves the performance.

Structure-Function Analysis of DNA Binding Domain of the Yeast ABF1 Protein (효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구)

  • Cho, Gi-Nam;Lee, Sang-Kyung;Kim, Hong-Tae;Kim, Ji-Young;Rho, Hyune-Mo;Jung, Gu-Hung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.102-108
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    • 1994
  • Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

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The amino acid analysis of polyhedrin and DNA sequence of ployhedrin gene in nuclear polyhedrosis virus (Nuclear polyhedrosis virus의 polyhedrin 아미노산 및 polyhedrin gene 염기서열 분석)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.37-46
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    • 1995
  • The amino acid analysis of polyhedrin protein and nucleotide sequence of polyhedrin gene in H. cunea nuclear polyhedrosis virus (HcNPV) genome have been studied. Polyhedrin had three polypeptide bands in SDS - polyactylamide gel electrophoresis. The major polypeptide had a molecular weight of 25 kd. The polyhedrin was composed of 17 different amino acids. HcNPV DNA was digested with EcoRI restriction enzyme and hybridized with ($\alpha^{32}P$) -labelled AcNPV polyhedrin gene cDNA. The polyhedrin gene was located on the fragment of EcoRI-H. The EcoRI - H fragment containing polyhedrin gene was cloned into the EcoRI site of pUC8 vector which was confirmed with southern blotting, and the recombinant plasmid containg polyhedrin gene was designated as hPE-H. The promoter region of polyhedrin genomic DNA was sequenced. The sequences identified as the TATA box was found at the 5' flanking region of the polyhedrin genomic DNA approximately -79 bp upstream from the transcriptional start site. But CAAT-like box was not shown near the TATA-like box in the polyhedrin gene. Four tandem repeats with the sequence 5' -CTAATAT-3' and 5'-TAAATAA-3' were found between -141 and -108 or -83 upstream and -52 bp downstream from the translation start site. About -141 bp region upstream from the translational start site was highly AT (78%) rich. The coding region for the polyhedrin starts and ends with ATG and TAA, respectively.

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