• 제목/요약/키워드: amino acid sequence analysis

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cDNA Cloning, Sequence Analysis and Molecular Modeling of a New Peptide from the Scorpion Buthotus saulcyi Venom

  • Nikkhah, Maryam;Naderi-Manesh, Hossein;Taghdir, Majid;Talebzadeh, Mehdi;Sadeghi-Zadeh, Majid;Schaller, Janatan;Sarbolouki, Mohamad N.
    • BMB Reports
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    • 제39권3호
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    • pp.284-291
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    • 2006
  • In this study, the cDNA of a new peptide from the venom of the scorpion, Buthotus saulcyi, was cloned and sequenced. It codes for a 64 residues peptide (Bsaul1) which shares high sequence similarity with depressant insect toxins of scorpions. The differences between them mainly appear in the loop1 which connects the $\beta$-strand1 to the $\alpha$-helix and seems to be functionally important in long chain scorpion neurotoxins. This loop is three amino acids longer in Bsaul1 compared to other depressant toxins. A comparative amino acid sequence analysis done on Bsaul1 and some of $\alpha$-, $\beta$-, excitatory and depressant toxins of scorpions showed that Bsaul1 contains all the residues which are highly conserved among long chain scorpion neurotoxins. Structural model of Bsaul1 was generated using Ts1 (a $\beta$-toxin that competes with the depressant insect toxins for binding to $Na^+$ channels) as template. According to the molecular model of Bsaul1, the folding of the polypeptide chain is being composed of an anti-parallel three-stranded $\beta$-sheet and a stretch of $\alpha$-helix, tightly bound by a set of four disulfide bridges. A striking similarity in the spatial arrangement of some critical residues was shown by superposition of the backbone conformation of Bsaul1 and Ts1.

Prediction of Rice Embryo Proteins using EST-Databases

  • Woo, Sun-Hee;Cho, Seung-Woo;Kim, Tae-Seon;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Jong, Seung-Keun
    • 한국육종학회지
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    • 제40권1호
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    • pp.1-7
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    • 2008
  • An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.

Random Sequence Analysis of the Genomic DNA of Methanopyrus kandleri and Molecular Cloning of the Gene Encoding a Homologue of the Catalytic Subunit of Carbon Monoxide Dehydrogenase

  • Shin, Hyun-Seock;Ryu, Jae-Ryeon;Han, Ye-Sun;Choi, Yong-Jin;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.404-413
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    • 1999
  • Methanopyrus kandleri is a hyperthermophilic methanogen that represents one of the most heat-resistant organisms: the maximum growth temperature of M. kandleri is $110^{\circ}C$. A random sequence analysis of the genomic DNA of M. kandleri has been performed to obtain genomic information. More than 200 unique sequence tags were obtained and compared with the sequences in the GenBank and PIR databases. About 30% of the analyzed tags showed strong sequence similarity to previously identified genes involved in various cellular processes such as biosynthesis, transport, methanogenesis, or metabolism. When statistics relating to the frequency of codons were examined, the sequenced open reading frames showed highly biased codon usage and a high content of charged amino acids. Among the identified genes, a homologue of the catalytic subunit of carbon monoxide dehydrogenase (CODH) that reduces $CO_2$ to CO was cloned and sequenced in order to examine its detailed gene structure. The cloned gene includes consensus promoters. The amino acid sequence of the cloned gene shows a strong homology with the CODH genes from methanogenic Archaea, especially in the presumed binding sites for Fe-S centers.

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

  • 권동현;변우현
    • 미생물학회지
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    • 제27권3호
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    • pp.194-200
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    • 1989
  • S. aureus에서 분리된 plasmid pSBK203 상의 CAT 유전자 염기서열을 결정하였으며 유발성 발현현상이 확인되었다. 염기서열 결과에 의해 예측된 단백질의 아미노산 서열 분석결고 pC221-CAT 와는 78%의 가장 높은 상동성을 나타냈으며 pC194-CAT와는 55%, 그람음성균 유래의 CAT 중 하나인 Tn9-CATdhkss 38%의 상동성을 각각 보여주고 있었다.

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Streptomyces somaliensis가 생산하는 세포외 Phospholipase D의 유전자 서열 분석과 Transphosphatidylation 활성 특성 (Nucleotide Sequence of an Extracellular Phospholipase D Gene from Streptomyces somaliensis and Transphosphatidylation Activity of Its Enzyme)

  • 정수진;이선희;엄태붕
    • 미생물학회지
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    • 제40권3호
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    • pp.211-216
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    • 2004
  • 세포외 phospholipase D (PLD)를 과량 생산하는 균주 JE-11을 토양으로부터 분리하였다. 16S rDNA에 의한 분석과 형태적, 생리적 특성을 조사한 결과 이 균은 Streptomyces somaliensis로 동정되었다. 선발한 S. somaliensis로 부터 PLD를 암호화하는 유전자(sspld) 분리하고 염기서열을 조사하였다. Open reading frame을 분석한 결과 33개의 아미노산으로 이루어진 분비 signal peptide와 505개의 아미노산으로 구성된 PLD단백질을 암호화하는 것으로 예상되었다. 또한, sspld의 염기 서열로부터 유추된 단백질 서열은 기존에 보고된 다른 Streptomyces PLD들과 70-88%의 서열 유사성을 보였다. 이 PLD는 96-98%(㏖/㏖)의 수율로서, Phosphatidylcholine을 glycerol과 serine을 기질로 하여 각각 phosphatidylglycerol 과phosphatidylserine으로 전환을 하였으나, 알코올 공여체인 inositol과 ethanolamine과는 반응하지 않았다.

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

  • 김도경;여상건
    • 대한수의학회지
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    • 제43권3호
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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.

RP-PCR을 이용한 보리누른모자이크바이러스 (BaYMV)와 보리마일드모자이크바이러스(BaMMV)의 외피단백질 유전자 검정 및 해석 (Analysis and Detection of Coast Protein Gene of Barley Yellow Mosaic Virus and Barley Mield Mosaic Virus by RT-PCR)

  • 이귀재
    • 한국식물병리학회지
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    • 제14권4호
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    • pp.314-318
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    • 1998
  • Using the reverse transcription polymerase chain reaction (RT-PCR), a rapid and sensitive assay method for the detection and identification of barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV) was adapted. Two units of primers from each virus were selected and used for the determination of two different viruses. PCR fragments of BaYMV (ca. 0.9kb) and BaMMV (ca. 0.8kb) were obtained from the designed method for the assay of BaYMV and BaMMV coat protein. PT-PCR fragments were cloned using vector pT7 Blue and the sequences of the selected clones were analyzed. coat protein of BaYMV and that of BaMMV consisted of 297 amino acids (891 nucleotides) and 251 amino acids (753 nucleotides), respectively. The snalysis of coat protein genes from these two viruses showed that 45.6% of nucleotides sequence ad 34.9% of amino acid in BaYMV were homologous to those in BaMMV.

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Isolation and Characterization of Exogenously Expressed Calmodulin from Endogenous Tobacco Calmodulin by Anion-exchange Fast Protein Liquid Chromatography

  • Oh, Suk-Heung;Cha, Youn-Soo;Lee, Tae-Kyoo
    • BMB Reports
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    • 제28권4호
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    • pp.306-310
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    • 1995
  • A Mono Q HR 5/5 anion-exchange column with a FPLC system was used to separate exogenously expressed calmodulin from endogenous tobacco calmodulins. Transgenic tobacco calmodulins were purified by fractionation with ammonium sulfate, precipitation with sulfuric acid and hydrophobic chromatography on phenyl-Sepharose CL-4B. The purified calmodulins were chromatographed in the FPLC using the column. This method was selected because of the slight differences in the net charge of foreign and endogenous plant calmodulins due to amino acid sequence differences. By this approach, the exogenously expressed calmodulin was isolated from endogenous tobacco calmodulins. The isolated calmodulin was characterized by amino acid composition analysis as well as methylation analysis.

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Bacillus subtilis subsp. krictiensis가 생산하는 항진균 물질 KRF-001의 구조 결정 (Structure Determination of Antifungal KRF-001 Produced by Bacillus subtilis subsp. krictiensis)

  • 김성기;이남경;정태숙;김영국;최진자;복성해
    • 한국미생물·생명공학회지
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    • 제19권6호
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    • pp.598-603
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    • 1991
  • 신규 미생물 Bacillus subtilis subsp. krictiensis의 배양액으로부터 식물 및 인체 병원균에 폭넓은 항진균 활성을 나타내는 DRF-001으로 총칭되는 6개의 cyclic peptide의 복합체(A에서 F)를 분리하였다. 이 6개 peptides의 분자량은 FAB-MS 측정결과, A가 1042, B와 C가 1056, D와 E가 1070 그리고 F가 1084였다. 이 복합체의 구조를 각종 자기분석을 통해 해석한 결과, 구조 중에 1몰씩의 glutamine, proline, tyrosine, serine 및 unusal $\beta$-amino acid와 3몰의 asparagine을 공통적으로 갖고 있으나, $\beta$-amino acid의 탄소수와 말단 methyl 구조에 차이점이 있음을 알 수있었다. 구성 amino acid들의 서열과 입체구조 결정에 의하여 KRF-001의 잠정구조를 결정하였다.

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Ribosomal Protein S4 Genes in Macaca fuscata: Sequence, Evolution, and Phylogeny

  • Kim, Heui-Soo
    • Journal of Life Science
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    • 제11권1호
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    • pp.34-38
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    • 2001
  • The cDNA encoding ribosomal protein S4(RPS 4) from an ovary cDNA library of the Japanese monkey (Macaca fuscata) was cloned and sequenced. The RPS4X gene from monkey X chromosome encodes a deduced protein of 263 amino acids and share 99.1% cDNA sequence similarity and 100% amino acid sequence identify with the human RPS4X. Rate of synonymous substitution was higher in RPS4Y than in RPS4X in comparison to the monkey and human. The ratio of synonymous and nonsynonymous substitutions per site indicated that directional selection has nor occurred in RPS4 genes. Phylogenetic analysis using the neighbor-joining method revealed that X and Y-linked RPS4 genes have evolved independently.

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