• 제목/요약/키워드: Amino acid sequence

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Molecular Characterization and Expression of CuZn-superoxide Dismutase (PSOD1) from Populus alba${\times}$Populus glandulosa

  • Lee Jun-Won;In Jun-Gyo;Lee Bum-Soo;Choi Yong-Eui;Kim Jin-Ju;Yang Deok-Chun
    • Plant Resources
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    • 제8권1호
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    • pp.52-59
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    • 2005
  • A cDNA, PSOD1, encoding cytosolic copper/zinc superoxide dismutase (CuZn-SOD) was cloned and characterized from a full length cDNA library prepared from Populus alba${\times}$Populus glandulosa cultured in vitro. A PSOD1, is 725 nucleotides long and has an open reading frame of 459 bp with 152 amino acid residues (pI 5.43). The deduced amino acid sequence of PSOD1 perfect matched to the previously reported CuZn-SOD (CAC33845.1). Consensus amino acid residues (His-45, -47, -62, -70, -79, -119) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of PSOD1 exhibited the high level of similarity from 100 to $85\%$ among previously registered SOD genes. The expression of PSOD1 in poplar increased at the 1 mM $H_{2}O_2$ and drought stress during 30 min and 60 min, but the ozone treated poplar increased at 30 min in the early time and then decreased at 60 min.

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Molecular Characterization of crp, the Cyclic AMP Receptor Protein Gene of Serratia marcescens KTCC 1272

  • Yoo, Ju-Soon;Kim, Hae-Sun;Chung, Soo-Yeol;Choi, Yong-Lark
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.670-676
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    • 2000
  • Several clones obtained from Serratia marcescens stimulated E. coli TP2139 (${\Delta}lac, \;{\Delta} crp$) cells to use maltose as a carbon source. The crp gene clone, pCKB12, was confirmed to stimulate the $\beta$-galactosidase activity, by Southern hybridization [31]. The nucleotide sequence of the crp region consisting of 1,979 bp was determined. The sequencing of the fragment led to the identification of two open reading frames: One of these, the crp gene, encoded 210 amino acid and the other encoded a truncated protein. The S. marcescens and E. coli crp genes showed a higher degree of divergence in their nucleotide sequence with 120 changes, however, the corresponding amino acid sequences showed only two amino acid differences. Yet, an analysis of the amino acid divergence revealed that the catabolite gene activator protein, the crp gene product, was the most conserved protein observed so far. Using a crp-lac protein fusion, it was demonstrated that S. marcescens CRP could repress its own expression, probably via a mechanism similar to that previously described for the E. coli crp gene.

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위치 종속 유사도 스펙트럼을 이용한 단백질 서열의 아미노산 조성 추정 (Estimating Amino Acid Composition of Protein Sequences Using Position-Dependent Similarity Spectrum)

  • 지상문
    • 한국정보과학회논문지:소프트웨어및응용
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    • 제37권1호
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    • pp.74-79
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    • 2010
  • 단백질의 아미노산 조성은 생물정보학의 여러 문제를 해결하기 위한 기초적인 정보로 자주 활용된다. 본 논문에서는 아미노산간의 진화적인 연관성을 정의한 BLOSUM 행렬에서 유도한 유사도 함수를 사용하여 아미노산 조성을 결정한다. 이러한 방법은 생물학적인 연관성이 있는 단백질 서열일수록 비슷한 아미노산 조성을 갖도록 한다. 또한 단백질의 구조와 기능에 중요한 역할을 하는 위치-특이적인 아미노산의 분포를 추정하기 위해서 레이더나 음성 신호의 스펙트럼 분석에 사용되는 개념인 시간-종속 분석, 시간 해상도와 주파수 해상도의 개념을 적용하였다. 제안한 방법을 단백질의 세포내 위치예측에 적용하여 기존의 아미노산 조성 추정 방법을 사용하는 것보다 크게 향상된 성능을 보임을 확인하였다.

Molecular Identification and Sequence Analysis of Coat Protein Gene of Ornithogalum mosaic virus Isolated from Iris Plant

  • Yoon, Hye-In;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제18권5호
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    • pp.251-258
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    • 2002
  • A potyvirus was isolated from cultivated Iris plants showing leaf streak mosaic symptom. Reverse transcription and polymerase chain reaction (RT-PCR) product of 1 kb long which encoded partial nuclear inclusion B and N-terminal region of viral coat protein (CP) genes for potyviruses was successfully amplified with a set of potyvirus-specific degenerate primers with viral RNA samples from the infected leaves: The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined. The nucleotide sequence of a CDNA clone revealed that the virus was an isolate of Ornithogalum moseic virus (OrMV) based on BLAST search analysis and was denoted as OrMV Korean isolate (OrMV-Ky). To further characterize the CP gene of the virus, a pair of OrMV-specific primers was designed and used for amplification of the entire CP gene of OrMV-Kr, The virus was easily and reliably detected from virus-infected Iris leaves by using the RT-PCR with the set of virus-specific primers. The RT-PCR product of the CP gene of the virus was cloned and its sequences were determined from selected recombinant CDNA clones. Sequence analysis revealed that the CP of OrMV-Kr consisted of 762 nucleotides, which encoded 253 amino acid residues. The CP of OrMV-Ky has 94.1-98.0% amino acid sequence identities (20 amino acid alterations) with that of other three isolates of OrMV, Two NT rich potential N-glycosylation motif sequences, NCTS and NWTM, and a DAC triple box responsible for aphid transmission were conserved in CPs of all the strains of OrMV. The virus has 58.5-86.2% amino acid sequence identities with that of other 16 potyviruses, indicating OrMV to be a distinct species of the genus. OrMV-Ky was the most related with Pterostylia virus Yin the phylogenetic tree analysis of CP at the amino acid level. This is the first report on the occurrence of OrMV in Iris plants in Korea. Data in this study indicate that OrMV is found in cultivated Iris plants, and may have mixed infection of OrMV and Iris severe mosaic virus in Korea.

Genomic Organization of ancop Gene for ${\alpha}-COP$ Homolog from Aspergillus nidulans

  • Lee, Hwan-Hee;Chae, Shun-Kee;Kim, Jeong-Yoon;Maeng, Pil-Jae;Park, Hee-Moon
    • Mycobiology
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    • 제28권4호
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    • pp.171-176
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    • 2000
  • We have cloned a ${\alpha}-COP$ homolog, ancop, from Aspergillus nidulans by colony hybridization of chromosome specific library using ${\alpha}-COP$ homologous fragment as a probe. The probe DNA was amplified with degenerated primers designed by comparison of conserved region of the amino acid sequences of Saccharomyces cerevisiae ${\alpha}-COP$, Homo sapiens HEP-COP, and Drosophila melanogaster ${\alpha}-COP$. Full length cDNA clone was also amplified by RT-PCR. Comparison of genomic DNA sequence with cDNA sequence obtained by RT-PCR revealed 7 introns. Amino acid sequence similarity search of the anCop with other ${\alpha}-COPs$ gave an overall identity of 52% with S. cerevisiae, 47% with human and bovine, 45% with Drosophila and Arabidopsis. In upstream region from the transcription start site, a putative TATA and CAAT motif were also identified.

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Nucleotide Sequence and Characterization of ptsG Gene Encoding Glucose-specific Enzyme II of Phosphotransferase System from Brevibacterium flavum

  • Yoon, Ki-Hong
    • Journal of Applied Biological Chemistry
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    • 제48권4호
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    • pp.218-221
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    • 2005
  • Nucleotide sequence of Brevibacterium flavum ptsG gene capable of complementing Escherichia coli ZSC113 mutations defective to glucose permease activity of phosphotransferase system was completely determined, and the gene product was compared with other glucose-specific enzyme II ($EII^{Glc}$). A ptsG gene of B. flavum consisted of open reading frame of 2,025 nucleotides putatively encoding polypeptide of 675 amino acid residues and TAA stop codon. Deduced amino acid sequence of B. flavum ($EII^{Glc}$) had high homology with ($EIIs^{Glc}$) of Corynebacterium glutamicum, C. efficiens, and B. lactofermentum. Arrangement of structural domains, IIBCA, of B. flanum ($EII^{Glc}$) protein was identical to that of EIIs belonging to glucose-phosphotransferase system.

Characterizations of Broad Benn Wilt Virus isolated from Rehmania glutinosa Liboschitz

  • Park, In-Sook;Park, Min-Kyoung;Kim, Jong-Yeop;Lim, Ju-Rak;Lee, Kui-Jae;Lee, Wang-Hyu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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    • pp.54-54
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    • 2002
  • The nucleotide sequence of a Korea isolate of broad bean wilt favavirus from Rehmannia glutinosa Lib., designated BBWV-RE, was determined. Direct amino acid sequencing of the virus coat proteins suggests that a comparison of several favaviruses in terms of nucleotide sequence and amino acid sequences showed that BBWV-2 isolates display high sequence identity. The small coat protein genes of RNA-2 were also determined for three other Japanese isolates(E, L, and 1-2) and two ATCC isolates(PV132 and PV176) of BBWV. the CP sequence suggested distinct evolution lineages. Serotype 2 favaviruses are more prevalent in Asia, Australia and North America, Wheres serotype 1 is more prevalent in europe.(중략)

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대두 $\beta$- Amylase의 N-말단 아미노산 배열 (N-Terminal Sequence of Soybean $\beta$- Amylase)

  • 지의상;김관묵;김준평
    • 한국식품영양학회지
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    • 제4권2호
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    • pp.161-166
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    • 1991
  • The blocked N-terminus and N-terminal sequence of soybean B-amylase were aetermined by analyzing the acidic peptides derived on peptic digestion of the enzyme. The acidic peptides were separated from the digest on a Dowex 50$\times$2 column(1X5cm) and purified by reversed phase-high performance liquid chromatography(RP-HPLC). The major acidic peptide, PEP-1, was a heptapeptlde. The N-terminal 7 amino acid sequence of soybean B-amylase was deduced to be acetyl-Ala-Thf-Ser-Asp-Ser-Asn-Met- from the results of sequence analysis of PEP-1 and amino acid analysis of other acidic peptides.

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국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현 (Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea)

  • 정동혁;송윤경;김경미;박효선;백명순;강신영
    • 대한수의학회지
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    • 제42권1호
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    • pp.89-100
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    • 2002
  • The nonstructural glycoprotein NSP4, encoded by the 10th gene of rotavirus, has been known to play important roles in viral assembly and pathogenesis. The NSP4 genes of human rotavirus Korean isolates, designated as CBNU/HR-1, CBNU/HR-2, CBNU/HR-3, and CBNU/HR-4, were cloned, sequenced and characterized. Also, the NSP4 gene of the CBNU/HR-1 was expressed in a baculovirus-insect cell system. The sequence data indicated that the NSP4 genes of human rotavirus Korean isolates were 750 or 751 bases in length and encoded one open reading frame of 175 amino acids. Two glycosylation sites were recognized in the NSP4 gene of human rotavirus isolates tested. The NSP4 of CBNU/HR-1, CBNU/HR-3, and CBNU/HR-4 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype B viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype A viruses. However, the NSP4 of CBNU/HR-2 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype A viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype B viruses. The Sf9 cells infected with recombinant baculovirus, inserted with NSP4 gene of CBNU/HR-1, produced specific cytopathic effects and the expressed NSP4 was detected by immunofluorescence staining using NSP4-specific monoclonal antibody(MAb). The expressed NSP4 migrated at 16-26 kDa on SDS-PAGE and reacted with NSP4-specific MAb by Western blotting.

Purification and Amino Acid Sequence of the Linoleate Isomerase Produced from Butyrivibrio fibrisolvens A-38

  • Park, Sook-Jahr;Park, Kyung-Ah;Park, Cjerl-Woo;Park, Won-Seck;Kim, Jeong-Ok;Ha, Yeong-Lae
    • Preventive Nutrition and Food Science
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    • 제1권2호
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    • pp.244-251
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    • 1996
  • Molecular weight and partial amino acid sequence of the cis, 9-cis, 12-octadecadienoate isomerase(linoleate isomerase) of Butyrivibrio fibrisovens A-38 were determined. Linoleate isomerase was isolated from the bac-teria cultured anaerobically and purified by ultracentrifugation in conjunction with Sepharose 6B column chro-matography, Phenyl sepharose 4B column chromatography and fast performance liquid chromatography (EPLC). The isomerase was single polypeptide with 19KD of molecular weight, when determined by SDS-PAGE. Fourteen amino acids sequence of N-terminal of the linoleate isomerase was N-GEIDKYPRIIKQQ determined by Edman method.

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