• Title/Summary/Keyword: amino acids sequence analysis

검색결과 390건 처리시간 0.021초

Cloning and DNA Sequence of Carboxymethylcellulase (CMCase) Gene from Cellulomonas sp. YE-5

  • Her, Song;Kim, Dong-Seob;Choi, Sun-Jin
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.86-90
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    • 1993
  • CMCase positive clones were screened from Cellulomonas sp. YE-5 and named pCE1, pCE2 and pCE3. Among the positive clones pCE1 was used for this study, because it has the smallest insert and the highest CMCase activity among the 3 clones, and its nucleotide sequence was determined. The CMCase gene in pCE1 was composed of 1071 bp of nucleotides coding 357 amino acids. Computer analysis showed that the pCE1 has 65% sequence homology with the endoglucanase from Cellulomonas fimi.

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dsRNA Analysis and Sequence of S12 to Rice dwarf virus Korean Isolate

  • Lee, Bong-Choon;Kwak, Do-Yeon;Hong, Yeon-Kyu;Cho, Hyun-Je;Park, Sung-Tae;Kim, Soon-Chul
    • The Plant Pathology Journal
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    • 제20권2호
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    • pp.155-157
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    • 2004
  • We isolated Rice dwarf virus (RDV) from infected plants in rice fields (Korea, Japan, China, the Philippines and Nepal) and analyzed their genomic dsRNAs by polyacrylamide gel eletrophoresis. The genomic dsRNAs of the isolates showed distinct electrophoretic mobility profiles. The S12 coding to nonstructural protein of Korean isolate (RDV-Kr) was further analyzed by sequencing. The S12 of RDV-Kr was 1,066bp long and coded for a protein composed of 312 amino acids including three open reading frames of P12, P120Pa and P120Pb. The sequence identities were 96% and 98.6% with Japanese isolates (H, AN), 94.7% with Nepalese isolate (NEL), 94% with Chinese isolate (CK) and the Philippines isolate (P).

Colletotrichum spp. 유래 scytalone dehydratase 유전자의 유전적 다양성 비교 (Genetic Variation of Scytalone Dehydratase Gene in Colletotrichum spp.)

  • 윤여홍;현민우;서동연;김용민;김성환;최창원;김영식
    • 한국균학회지
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    • 제37권2호
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    • pp.155-160
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    • 2009
  • The production of dihydroxynaphthalene (DHN) melanin is known to be essential factor for pathogenicity in Colletotrichum lagenarium. However, the genetic diversity of melanin genes was not much known among Colletotrichum spp. To investigate the variability of melanin gene in Colletotrichum spp. that cause anthracnose on diverse crops including tomato, we cloned and sequenced partial sd, one of DHN melanin genes encoding for scytalone dehydratase, from eight strains of C. coccodes, C. acutatum, C. truncatum C. caricae, and C. musae. The size of PCR-amplified sd ranged 437 bp to 545 bp. The nucleotide sequence identity of sd among the Colletotrichum strains tested varied from 49% to 99%. All of the PCR-amplified sd from eight strains contain an intron and have two exons coding for 122 amino acids. Overall, the size and nucleotide sequence of sd varied among the five Colletotrichum spp. Sequence identity of the predicted scytalone dehydratase protein of 122 amino acids ranged 50 to 99%. Phylogentic analysis based on the sd nucleotide sequences revealed that the five Colletotrichum spp. could be genetically divided.

Protein Hydrolysis with Formic Acid and Analysis of Amino Acid Using Butylthiocarbamyl - trimethylsilyl (BTC - TMS) Derivatives by Gas Chromatography

  • 우강융;이동선;김민철
    • 한국식품영양과학회지
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    • 제32권3호
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    • pp.320-324
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    • 2003
  • 단백질 구성 아미노산 분석을 위한 효과적인 가수분해 방법을 찾기 위하여 0.3% tryptamine을 함유한 6M formic acid와 6M HCI을 표준 아미노산과 단백질 표준품인 bovine serumn albumin 가수분해에 적용하여 표준아미노산의 회수율과 bovine serum albumin의 아미노산 조성을 분석하였고 GC에 의한 효과적인 아미노산 분석을 위하여 새로운 유도체인 butylthiocarbamyl-trimethylsilyl(BTC-TMS)유도체를 개발하여 분석한 결과는 다음과 같았다 표준아미노산의 회수율은 6M formic acid에 의한 가수분해방법이 6M HCI에 의한 가수분해 방법보다 상당히 정확하였고 특히 산 가수분해에서 tryptamine의 존재하에서도 잘 파괴되는 tryptophan의 경우 formic acid가수분해가 HCI가수분해보다 1.5배정도 높은 회수율을 보였다. Bovine serum albumin의 아미노산 조성을 583 아미노산 잔기로 환산하여 나타내었을 경우도 formic acid에 의한 가수분해가 HCI가수분해 경우보다 훨씬 정확하였고 이 때 tryptophan의 회수율도 훨씬 높게 나타났다. 다만 서열분석에서 분석되지 않는 cystine이 formic acid 가수분해시 분석되고 있어 이에 대한 정확한 검정이 필요하였다. BTC-TMS 유도체는 GC분석시 극성이 다소 낮은 DB-l7 column으로 분리가 잘되었고 재현성도 좋았으나 GC분석을 위한 대부분의 유도체에서와 마찬가지로 몇 가지 아미노산에서 두 개의 peak로 나타나는 결점이 있었다.

Isolation and Characterization of a cDNA for a Ribulose-1,5-Bisphosphate Carboxylase Small Subunit in Spinach

  • Jin, Yun-Hae;Park, Yang-Seo;Jeong, Ji-Na;Cho, Tae-Ju;Cho, Nam-Jeong
    • BMB Reports
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    • 제30권3호
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    • pp.173-176
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    • 1997
  • We isolated a cDNA clone that encodes a ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) from spinach using a soybean rbcS cDNA as a probe. The small subunit consists of 180 amino acids including a transit peptide of 57 residues. Comparison of the amino acid sequence with those of other plant species shows a maximum of 70-80% identical residues. Southern blot analysis suggests the existence of multiple rbcS genes in the spinach genome. Northern blot analysis indicates that the rbcS gene is expressed predominantly in leaves and that the expression of the gene is induced by light.

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A cDNA Clone for the 5' Exon of Chloroplast ATP Synthase Subunit I Gene (atpF) from Broccoli (Brassica oleracea L. var. Italica) and Its Expression Pattern

  • Choo Bong Hong
    • Journal of Plant Biology
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    • 제38권2호
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    • pp.137-141
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    • 1995
  • We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

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Molecular Cloning, Gene Structure, Expression, and Enzyme Activity of a Serine Protease from Water Scorpion, Laccotrephes japonensis (Hemiptera: Nepidae)

  • Park, Kwan Ho;Choi, Young Cheol;Nam, Seong Hee;Hwang, Jae Sam;Nho, Si Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제25권2호
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    • pp.187-193
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    • 2012
  • Serine proteases are major insect enzymes involved in the digestion of dietary proteins and in the process of blood meal digestion. In this study, cDNA was constructed using the whole body of Laccotrephes japonensis. The flanking sequences of the 5- and 3- end of this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained a 963-bp ORF encoding 320 amino acids. The deduced amino acid sequence showed 62% identity with the Creontiades dilutus serine protease, 58% with the Lygus lineolaris trypsin precursor, and 54% with the Triatoma infestans salivary trypsin. To assess the expression of the L. japonensis serine protease (JGsp), the JGsp gene was cloned into a baculovirus transfer vector, pBac-1, and expressed in Sf9 cells (Spodoptera frugiperda). SDS-PAGE and western blot analysis have shown that the JGsp recombinant protein was a monomer with a molecular weight of about 32 kDa. Recombinant JGsp has shown activity in the protease enzyme assay using gelatin as a substrate.

A Myostain-like Gene Expressed Highly in the Muscle Tissue of Chinese mitten crab, Eriocheir sinensis

  • Kim, Kyoung-Sun;Jeon, Jeong-Min;Kim, Hyun-Woo
    • Fisheries and Aquatic Sciences
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    • 제12권3호
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    • pp.185-193
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    • 2009
  • A complete cDNA, which encodes for a myostatin-like protein (Es-MSTN), was isolated from the Chinese mitten crab, Eriocheir sinensis. Es-MSTN was composed of 2,397 nucleotides and the open reading frame (ORF) specified a protein containing 468 amino acids. Es-MSTN exhibited 32% amino acid sequence identity and 52% similarity to human myostatin. Multiple sequence alignment analysis indicated that Es-MSTN possessed the conserved proteolytic cleavage site (RXXR) for maturation of the protein and nine cysteine residues for disulfide bridges. Besides the conserved structural features, Es-MSTN also exhibits its unique characters; a longer N-terminal domain which is involved in protein folding and latent form of myostatin and absence of the cleavage site for BMP-1/tolloid family of metalloproteinase to activate mature myostatin. Phylogenetic analysis suggests that Es-MSTN showed the closely related to both vertebrate myostatin and GDF11. Es-MSTN is expressed highly in the claw muscle, leg muscle, thoracic muscle and heart, and moderately in the hindgut suggesting that Es-MSTN may play important roles in the muscle tissues. As homolog of mammalian myostatin and GDF11, Es-MSTN may be involved in development of muscular tissue and further study will help to produce high-quality seafood.

Identification of Novel Esterase from Metagenomic Library of Yangtze River

  • Wu, Chao;Sun, Baolin
    • Journal of Microbiology and Biotechnology
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    • 제19권2호
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    • pp.187-193
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    • 2009
  • A metagenomic library of surface-water microbes from the Yangtze River in China was constructed, and a novel esterase, designated as EstY, was isolated and characterized. EstY had 423 amino acids with an estimated molecular mass of 44 kDa and pI of 7.28. It hydrolyzed various p-nitrophenyl esters(acetate, butyrate, caprate, caprylate, laurate, myristate, and palmitate) and its best substrate was p-nitrophenyl caprate(C8). The optimum pH for EstY activity was 9.0 and the optimum temperature was $50^{\circ}C$. Metal ions, such as $Mn^{2+},\;Co^{2+},\;Hg^{2+},\;Zn^{2+},\;and\;Fe^{3+}$, strongly inhibited the activity of EstY, whereas $Mg^{2+}$ was required for maximal activity. Activity remained in the presence of 10% alcohol, acetone, isopropanol, and dimethyl sulfoxide, respectively. An analysis of the amino acid sequence deduced from estY revealed that it had 7 closely related lipolytic enzymes. Moreover, a sequence analysis showed that EstY, like its 7 relatives, did not belong to any known lipolytic enzyme family.

최근 국내 분리 고병원성 infectious bursal disease virus의 segment A 유전자 특성 (Sequence analysis of segment A gene of a very virulent infectious bursal disease virus recently isolated in Korea)

  • 오현석;이진화;권혁무;성환우
    • 대한수의학회지
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    • 제51권1호
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    • pp.37-46
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    • 2011
  • Infectious bursal disease virus (IBDV) is a member of the Avibirnavirus genus of the Birnaviridae family which genome consists of two segments (A and B) of double stranded RNA. Segment A gene of KNU08010 isolate, which was isolated from a 15-day-old chicken flock in 2008, was sequenced and compared with other IBDV isolates including SH/92 strain, the first Korean very virulent (vv) IBDV isolate. The amino acid sequences of segment A gene showed that KNU08010 had 99.2% homology with SH92 strain. KNU08010 isolate had specific amino acids A222, I242, I256, I294 and S299 which are highly conserved among vvIBDV strains. Phylogenetic analysis based on the nucleotide sequences of variable region of the VP2 gene of 18 IBDV strains revealed that KNU08010 was grouped with vvIBDVs and was closely related to Korean vvIBDVs isolated from wild birds.