• 제목/요약/키워드: Open Reading Frame

검색결과 700건 처리시간 0.027초

Brevibacterium ammoniagenes의 DNA Polymerase I 유사 유전자의 분석 (Analysis of a Putative DNA Polymerase I gene in Brevibacterium ammoniagenes.)

  • 오영필;윤기홍
    • 한국미생물·생명공학회지
    • /
    • 제30권2호
    • /
    • pp.105-110
    • /
    • 2002
  • The sequence of 3,221 nucleotides immediately adjacent to rpsA gene encoding 30S ribosomal protein S1 of Brevibacterium ammoniagenes was determined. A putative open reading frame (ORF) of 2,670 nucleotides for a polypeptide of 889 amino acid residues and a TAG stop codon was found, which is located at a distance of 723 nucleotides upstream from rpsA gene with same translational direction. The deduced amino acid sequence of the ORF was found to be highly homologous to the DNA polymerase I of Streptomyces griseus (75.48%), Rhodococcus sp. ATCC 15963 (56.69%), Mycobacterium tuberculosis (55.46%) and Mycobacterium leprae (53.99%). It was suggested that the predicted product of the ORF is a DNA polymerase I with three functional domains. Two domains of 5 → 3 exonuclease and DNA polymerase are highly conserved with other DNA polymerase I, but 3 → 5 exonuclease domain is less conserved.

Cloning and Sequencing of the ddh Gene involved in the Novel Pathway of Lysine Biosynthesis from Brevibacterium lactofermentum

  • Kim, Ok-Mi;Kim, Hyun-Jeong;Kim, Dal-Sang;Park, Dong-Chul;Lee, Kap-Rang
    • Journal of Microbiology and Biotechnology
    • /
    • 제5권5호
    • /
    • pp.250-256
    • /
    • 1995
  • The ddh gene encoding meso-diaminopimelate (meso-DAP)-dehydrogenase (DDH) in Brevibacterium lactofermentum was isolated by complementation of the Escherichia coli dapD mutation. It was supposed from subcloning experiments and complementation tests that the evidence for DDH activity appeared in about 2.5 kb Xhol fragmented genome. The 2.5 kb Xhol fragment containing the ddh gene was sequenced, and an open reading frame of 960 bp encoding a polypeptide comprising 320 amino acids was found. Computer analysis indicated that the deduced amino acid of the B. lactofermentum ddh gene showed a high homology with that of the Corynebacterium glutamicum ddh gene.

  • PDF

Gene Cloning, High-Level Expression, and Characterization of an Alkaline and Thermostable Lipase from Trichosporon coremiiforme V3

  • Wang, Jian-Rong;Li, Yang-Yuan;Liu, Danni
    • Journal of Microbiology and Biotechnology
    • /
    • 제25권6호
    • /
    • pp.845-855
    • /
    • 2015
  • The present study describes the gene cloning and high-level expression of an alkaline and thermostable lipase gene from Trichosporon coremiiforme V3. Nucleotide analysis revealed that this lipase gene has an open reading frame of 1,692 bp without any introns, encoding a protein of 563 amino acid residues. The lipase gene without its signal sequence was cloned into plasmid pPICZαA and overexpressed in Pichia pastoris X33. The maximum lipase activity of recombinant lipase was 5,000 U/ml, which was obtained in fed-batch cultivation after 168 h induction with methanol in a 50 L bioreactor. The purified lipase showed high temperature tolerance, and being stable at 60℃ and kept 45% enzyme activity after 1 h incubation at 70℃. The stability, effects of metal ions and other reagents were also determined. The chain length specificity of the recombinant lipase showed high activity toward triolein (C18:1) and tripalmitin (C16:0).

Cloning of the Transketolase Gene from Erythritol-Producing Yeast Candida magnoliae

  • Yoo, Boung-Hyuk;Park, Eun-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • 제24권10호
    • /
    • pp.1389-1396
    • /
    • 2014
  • The entire nucleotide sequence of the TKL1 gene encoding transketolase (TKL) in an erythritol-producing yeast of Candida magnoliae was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed an open reading frame of C. magnoliae TKL1 (CmTKL1) that spans 2,088 bp and encodes 696 amino acids, sharing 61.7% amino acid identity to Kluyveromyces lactis TKL. Functional analysis showed that CmTKL1 complemented a Saccharomyces cerevisiae tkl1 tkl2 double mutant for growth in the absence of aromatic amino acids and restored transketolase activity in this mutant. An enzyme activity assay and RT-PCR revealed that the expression of CmTKL1 is induced by fructose, $H_2O_2$, and KCl. The GenBank accession number for C. magnoliae TKL1 is KF751756.

고려인삼으로부터 Peroxidase 유전자의 Cloning 및 Activity

  • 손화;심주선;양덕춘
    • 고려인삼학회:학술대회논문집
    • /
    • 고려인삼학회 2004년도 추계 학술대회 및 정기총회
    • /
    • pp.48-49
    • /
    • 2004
  • A peroxidase[E.C.1.11.1.7] is very important enzymes,e.g., as preventive antioxidants. The function is connected with growth and specialization of plant. It makes from the peroxidase and other product to save itself When a plant have been under stress of environment. A class III peroxidase cDNA was isolated from the flower bud of Panax ginseng C.A. Meyer and named PgPrx3. The PgPrx3 is an ORF(open reading frame) of 1,065 bp and a amino acid of 355 residue. Used BioEdit software to compare the PgPrx3 amino acid sequence with other plants which have already known a result of Identity was Spinacia oleracea(70%), vigna angularis(71%), Nicotiana tabacum(69%) and Linum usitatissimym(65%). The peroxidase of Vigna angularis has high homology relationship with ginseng. for that reason, the PgPx3 is a member of class III peroxidase.

  • PDF

cDNA Cloning of a Putative Alcohol Dehydrogenase from the Silkworm, Bombyx mori

  • Kim, Iksoo;Park, Yong-Soo;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제7권1호
    • /
    • pp.51-57
    • /
    • 2003
  • A cDNA encoding a putative alcohol dehydrogenase (ADH) class III was cloned from the silkworm, Bombyx mono The full length cDNA is 1,385 nucleotides long and contains an open reading frame of 1,128 bp encoding 376 amino acid residues. The B. mon ADH III protein sequence was aligned with ADH III known from various organisms. Interestingly, the protein sequence of B. mon ADH III showed 87% and 85% identity to ADH III from marine fish Sparus aurata and Branchiostoma floridae, respectively, whereas rather low sequence identity (83%) to Drosophila melanogaster ADH III was observed. Northern blot analysis revealed that B. mon ADH III mRNA is expressed in all tissues from larva examined: fat body, midgut, epidermis, silk gland and ovary, with the highest level found in the fat body.

Identification and Characterization of Bombyx mori LDH Gene through Bioinformatics Approaches

  • Zhu, Minfeng;Chen, Keping;Yao, Qin
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제15권2호
    • /
    • pp.137-143
    • /
    • 2007
  • Lactate dehydrogenase (LDH) is a ubiquitous enzyme that plays a significant role in the clinical diagnosis of pathologic processes. Discovery of the LDH (BmLDH) gene in B. mori may shed light on its role in the biology of Lepidoptera species, and afford further understanding of the function of the enzyme. In this study, we used the bioinformatics tools to identify LDH gene in B. mori. Sequence analysis showed that BmLDH cDNA contains a 996 bp open reading frame, encoding 331 AA proteins, with seven introns. Compared with hHLDH (human heart LDH), BmLDH contained the same key active sites. Domain search and protein fold recognition analyses provide compelling evidences that the deduced protein is a LDH. Using the computer program MEGA3, we conducted a search for homologs of BmLDH among many eukaryotic species and confirmed that the BmLDH was conserved in all organisms investigated. This gene has been registered in GenBank under the accession number EU000385.

Cloning and expression of glutathione S-transferase (GST) cDNA from Gossypium hirsutum L.

  • Kang, Won-Hee;Kim, Myong-Jo;Lim, Jung-Dae;Yun, Song-Joong;Chung, Ill-Min;Yu, Chang-Yeon
    • 한국약용작물학회지
    • /
    • 제10권4호
    • /
    • pp.294-297
    • /
    • 2002
  • A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.

Improvement in the Stability of Glycinecin A through Protein Fusion of the Two Structural Components

  • Kim, Youngmee;Somi K. Cho;Moonjae Cho
    • Journal of Microbiology
    • /
    • 제39권3호
    • /
    • pp.177-180
    • /
    • 2001
  • Glycinecin A, a bacteriocin produced by Xanthomonas campestris pv. glycines, inhibits the growth of X. c. pv. vesicatoria. We have reported that purified glycinecin A is composed of two polypeptides, is active over a wide range of pH (6 to 9), and is stable at temperatures up to 60$\^{C}$. Glycinecin A is a heterodimer consisting of 39- and 14-kDa subunits; the two encoding genes, glyA and glyB, respectively, have been cloned (Heu et al. 2001. Appl. Environ. Microbiol. 67, 4105-4110). Co-expression of glyA and glyB in the same cell is essential for bacteriocin activity. We constructed and produced a chimeric glycinecin A connecting glyA and glyB in one open reading frame. The chimeric glycinecin A has the same bactericidal activity as the wild-type glycinecin A. However, the chimeric glycinecin A is more stable in a wider range of pH and temperature.

  • PDF

지능적 다중염기서열 변환 도구의 설계 및 구현 (Design and Implementation of an Intelligent Multiple DNA Sequence Translation Tool)

  • 이혜리;이건명;이찬희;이성덕;김성수
    • 한국지능시스템학회:학술대회논문집
    • /
    • 한국퍼지및지능시스템학회 2006년도 춘계학술대회 학술발표 논문집 제16권 제1호
    • /
    • pp.37-40
    • /
    • 2006
  • 계통분석을 하는 생물학자들은 관련된 분석대상에 대한 정보를 확보하여 비교분석하기 위해 NCBI 등으로부터 염기서열을 확보하여 아미노산 서열로 변환하는 작업을 수행하게 된다. 많은 서열 데이터에 대해서 데이터베이스로부터 데이터를 검색하고 이를 변환하는 작업을 순차적으로 분석자가 관여하여 작업하는 것이 현재 분석환경이다. 따라서 본 논문에서는 분석의 효율성을 향상시키기 위해, 관심서열의 등록번호(Accession Number) 리스트를 입력하면 해당 서열에 대항 정보를 NCBI로부터 웹로봇을 통해 자동으로 확보한 다음, 확보된 염기서열 전체를 아미노산 서열로 자동 변환하여 가장 긴 ORF(Open Reading Frame)을 추천해주기 위해 설계된 지능형 다중 염기서열 변환 도구에 대해서 소개한다.

  • PDF