• Title/Summary/Keyword: Nucleotide sequence

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Molecular Cloning of the nahC Gene Encoding 1,2-Dihydroxynaphthalene Dioxygenase from Pseudomonas fluorescens

  • KIM, YEO-JUNG;NA-RI LEE;SOON-YOUNG CHOI;KYUNG-HEE MIN
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.172-175
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    • 2002
  • The complete nucleotide sequence of the nahC gene from Pseudomonas fluorescens, the structural gene for 1,2-dihydroxynaphthalene (1,2-DHN) dioxygenase, was determined. The 1,2-DHN dioxygenase is an extradiol ring-cleavage enzyme that cleaves the first ring of 1,2-dihydroxynaphthalene. The amino acid sequence of the dioxygenase deduced from the nucleotide sequence suggested that the holoenzyme consists of eight identical subunits with a molecular weight of approximately 34,200. The amino acid sequence of 1,2-DHN dioxygenase showed more than $90\%$ homology with those of the dioxygenases of other Pseudomonas strains. However, sequence similarity with those of the Sphingomonas species was less than $60\%$. The nahC gene of P. fluorescens was moderately expressed in E. coli NM522, as determined by enzymatic activity.

Trend and Technology of Gene and Genome Research (유전자 및 유전체 연구 기술과 동향)

  • 이진성;김기환;서동상;강석우;황재삼
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.126-141
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    • 2000
  • A major step towards understanding of the genetic basis of an organism is the complete sequence determination of all genes in target genome. The nucleotide sequence encoded in the genome contains the information that specifies the amino acid sequence of every protein and functional RNA molecule. In principle, it will be possible to identify every protein resposible for the structure and function of the body of the target organism. The pattern of expression in different cell types will specify where and when each protein is used. The amino acid sequence of the proteins encoded by each gene will be derived from the conceptional translation of the nucleotide sequence. Comparison of these sequences with those of known proteins, whose sequences are sorted in database, will suggest an approximate function for many proteins. This mini review describes the development of new sequencing methods and the optimization of sequencing strategies for whole genome, various cDNA and genomic analysis.

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Nucleotide Sequence Homology in Rotaviruses (Rotaviruses의 염기배열 유사성 측정)

  • ;Spendlove, Rex S.;Barnett, Bill B
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.155-161
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    • 1988
  • Nucleotide sequence homology between bovine, simian, and porcine rotavirus was determined by the RNA:RNA hybridization technique. Single stranded RNA, prepared in vitro with EDTA activated endogeneous viral RNA polymerase, was hhbridized with tritium labeled bovine rotavirus genomic RNA. The heteroduplex RNA was treated with single stranded RNA specific ribonucleases and the RNase resistant hybrid RNA was precipitated, and collected by filtration on a filter paper. Seventy four percent RNA sequence homology between bovine and simian rotavirus and 8 percent RNA sequence homology between bovine and porcine rotavirus was confirmed by hybridization between tritium labeled single stranded RNA and viral genomic RNA.

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Sequence Analysis of NS4 Region of HCV Isolated from Korean Patient

  • Paik, Sang-Hoon;Lee, Young-Ik;Kim, Won-Bae;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.260-266
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    • 1995
  • Hepatitis C virus (HCV) has been considered as a mojor causative agent of post-transfusion related non-A, non-B hepatitis. In this study, the cDNA sequence of NS4 region of HCV (HCV-S) obtained from a Korean patient's plasms was determined. Comparative nucleotide sequence analysis between to type II. 67.2% homology to type III, and 66.4% homology to type IV. The putative amino acid sequence homologies to types I, II, III, and IV were 82.8-84.7%, 92.5-95.1%. 72.5, and 71.1%, respectively. This data strongly suggests that HCV-S should be classified as type II. Significant similarities of hydrophobicity profiles and putative transmembranous domains were found in HCV-S and four major prototypes, indicating that the protein structure is similar in spite of the heterogeneities of intertype homologies at the level of the psrimary nucleotide and amino acid sequences.

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Analysis of Nucleotide Sequence Encoding VP2 Protein of Infectious Bursal Disease Virus Detected in Korea (국내 분리 닭 전염성 F낭병 바이러스의 VP2 단백질 생산 유전자의 염기서열 분석)

  • Kim, Toh-kyung;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.439-448
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    • 2003
  • The VP2 gene of infectious bursal disease virus (IBDV) Chinju which was previously detected in Chinju, Korea was cloned and sequenced to establish the information for the development of genetically engineered vaccines and diagnostic reagents against IBDV. The nucleotide sequence of the entire Chinju VP2 gene consisted of 1,356 bases long encoding 452 amino acids in a single open reading frame (ORF). It consisted of 368 adenine (27.1%), 363 cytosine (26.8%), 339 guanine (25.0%) and 286 thymine (21.1%) residues. The predicted $M_r$ of the Chinju VP2 protein was 48 kDa, and the protein contained 13 phosphorylation sites by protein kinase C, casein kinase II or tyrosine kinase, whereas 3 asparagine-linked glycosylation sites were recognized. The nucleotide sequence of Chinju VP2 ORF had a very close phylogenetic relationship with 98-99% homology to that of the very virulent IBDVs (vvIBDVs) HK46, OKYM, D6948, UK661, UPM97/61 and BD3/99. Also, the Chinju VP2 protein revealed a very close phylogenetic relationship with 99-100% homology to that of these vvIBDVs. The Chinju VP2 protein had 100% amino acid identity in the variable region of residues 206-360 with that of the D6948, HK46, OKYM and UK661, as well as 100% identity in two hypervariable regions of residues 212-224 and 314-324 with those of the D6948, HK46, OKYM, UK661, UPM97/61 and BD3/99. The amino acid sequence of the chinju VP2 protein contained a serine-rich heptapeptide of SWSASGS as in these vvIBDVs.

Implementation of an Information Management System for Nucleotide Sequences based on BSML using Active Trigger Rules (BSML 기반 능동 트리거 규칙을 이용한 염기서열정보관리시스템의 구현)

  • Park Sung Hee;Jung Kwang Su;Ryu Keun Ho
    • Journal of KIISE:Databases
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    • v.32 no.1
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    • pp.24-42
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    • 2005
  • Characteristics of biological data including genome sequences are heterogeneous and various. Although the need of management systems for genome sequencing which should reflect biological characteristics has been raised, most current biological databases provide restricted function as repositories for biological data. Therefore, this paper describes a management system of nucleotide sequences at the level of biological laboratories. It includes format transformation, editing, storing and retrieval for collected nucleotide sequences from public databases, and handles sequence produced by experiments. It uses BSML based on XML as a common format in order to extract data fields and transfer heterogeneous sequence formats. To manage sequences and their changes, version management system for originated DNA is required so as to detect transformed new sequencing appearance and trigger database update. Our experimental results show that applying active trigger rules to manage changes of sequences can automatically store changes of sequences into databases.

Nucleotide Sequence of Cellulolytic Xylanase Gene (bglBC2) from Bacillus circulans (Bacillus circulans 유래 cellulolytic xylanase 유전자(bglBC2)의 염기서열 결정 및 분석)

  • Kim, Ji-Yeon
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.67-72
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    • 2006
  • The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.

Characterization of the pcbE Gene Encoding 2-Hydroxypenta-2,4-Dienoate Hydratase in Pseudomonas sp. DJ-12

  • Lim, Jong-Chul;Lee, Jeongrai;Jang, Jeong-Duk;Lim, Jai-Yun;Min, Kyung-Rak;Kim, Chi-Kyung;Kim, Young-Soo
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.187-195
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    • 2000
  • Nucleotide sequence extending 2,3-dihydroxybiphenyl 1,2-dioxygenase gene (pcbC) and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase gene (pcbD) of Pseudomonas sp. DJ-12 was previously analyzed and the two genes were present in the order of pcbD-pcbC preceded by a promoter from Pseudomonas sp. DJ-12. In this study, a 3.8-kb nucleotide sequence located downstream of the pcbC gene was analyzed to have three open reading frames (ORFs) that are designated as orf1, pcbE and orf2 genes. All of the ORFs were preceded by each ribosome-binding sequence of 5-GGAXA-3 (X=G or A). However, no promoter-like sequence and transcription terminator sequence were found in the analyzed region, downstream of pcbC gene. Therefore, the gene cluster appeared to be present in the order of pcbD-pcbC-orf1-pcbE-orf2 as an operon, which is unique organization characterized so far in biphenyl- and PCB-degrading bacteria. The orf1 gene was composed of 1,224 base pairs which can encode a polypeptide of molecular weight 44,950 containing 405 amino acid residues. A deduced amino acid sequence of the orf1 gene product exhibited 21-33% identity with those of indole dioxygenase and phenol hydroxylase components. The pcbE gene was composed of 783 base pairs encoding 2-hydroxypenta-2,4-dienoate hydratase involved in the 4-chlorobiphenyl catabolism. The orf2 gene was composed of 1,017 base pairs encoding a polypeptide of molecular weight 37,378 containing 338 amino acid residues. A deduced amino acid sequence of the orf2 gene product exhibited 31% identity with that of a nitrilotriacetate monooxygenase component.

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Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203 (Staphylococcus aureus에서 분리된 R-plasmid pSBK203상의 chloramphenicol acetyltransferase 인자의 염기서열 및 유발성 분석)

  • 권동현;변우현
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.194-200
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    • 1989
  • The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.

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Sequence Analysis of the Coat Protein Gene of Citrus Tristeza Virus Isolated form Cheju Island

  • Park, Hyoun-Hyang;Kim, Dae-Hyun;Hyun, Woo-Taek;Moon, Doo-Khil;Koh, Young-jin;Park, Tae-Jin
    • The Plant Pathology Journal
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    • v.16 no.1
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    • pp.43-47
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    • 2000
  • Citrus tristeza virus (CTV) is the causal agent of one of the most important diseases of citrus. Recently, CTV has been detected in Cheju Island by ELISA. The coat protein (CP) gene of CTV isolated form Cheju Island was cloned by RT-PCR and the nucleotide was analyzed in this study. Citrus leaves were collected from trees showing decline symptoms from various region of Cheju Island in the summer of 1998 and 1999. The CP gene open reading frame is composed of 670 nucleotides and encodes a polypeptide of 223 amono acids. Sequence analysis the CP gene revealed that two CTV strains present in Cheju Island. Viruses collected form Sogwipo area and Cheju City area in 1999 ahowed 91-93% nucleotide sequence homology with CTV T36 strain. Viruses collected form Cheju City area in 1999 and Sogwipo City in 1998 showed 94-98% nucleotide sequence homology with CTV SY568 strain. A efficient viral RNA extraction methods was developed by modifying procedure for animal virus RNA purification methods and PCR product was detected form one tenth of RNA purified from as small as 45 mg fresh or frozen tissue.

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