• Title/Summary/Keyword: DNA sequence alignment

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Cloning and Characterization of the Putative Transferrin Receptor cDNA from the Olive Flounder (Paralichthys olivaceus)

  • Won Kyoung-Mi;Park Soo-Il
    • Fisheries and Aquatic Sciences
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    • v.6 no.2
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    • pp.101-104
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    • 2003
  • A cDNA clone for the olive flounder (Paralichthys olivaceus) transferrin receptor (fTfR) was isolated from a leukocytes cDNA library. The fTfR gene consisted of 2,319 bp encoding 773 amino acid residues. The amino acid sequence alignment of the fTfR showed that their size and hydrophobic profile are similar. In addition, the Tyr-Thr-Arg-Phe (YTRF) motif that is the recognition signal for high-efficiency endocytosis, is conserved very well. This motif is important for functional properties of TfR. The deduced amino acid sequence had $42.4-42.9\%$ identities with the previously reported TfRs of vertebrates. The fTfR was expressed in the blood, kidney, spleen, and liver of healthy olive flounder by the Northern blot hybridization.

Isolation and Characterization of Two Amino Acid-activating Domains of Peptide Synthetase Gene from Bacillus subtilis 713

  • Lee, Youl-Soon;You, Sang-Bae;Lee, Ji-Wan;Kim, Tae-Young;Kim, Sung-Uk;Bok, Song-Hae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.399-405
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    • 1998
  • From the sequence alignment of various non-ribosomal peptide synthetases, several motifs of highly conserved sequences have been identified within each domain of peptide synthetases. We designed PCR primers based on the highly conserved nucleotide sequences to amplify and isolate a ∼7.2-kb DNA fragment of the Bacillus subtilis 713 which was isolated and reported to produce an antifungal peptide compound. Nucleotide sequence analysis of 4.8 kb of the predicted amino acids revealed significant homology to various peptide synthetases over the whole sequence and also revealed two amino acid-activating domains with highly conserved Core 1 to Core 6 and spacer motif. This suggests that the isolated DNA fragment is part of a peptide synthetase gene for antifungal peptide.

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SNPchaser : A Web-based Program for Detecting SNPs Substitution and Heterozygosity Existence (SNPchaser : DNA서열의 SNPs 치환 및 Heterozygosity 확인 프로그램)

  • Jang, Jin-Woo;Lee, Hyun-Chul;Lee, Myung-Hoon;Choi, Yeon-Shik;Choo, Dong-Won;Park, Kie-Jung;Lee, Dae-Sang
    • KSBB Journal
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    • v.24 no.4
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    • pp.410-414
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    • 2009
  • Single-nucleotide polymorphisms (SNPs) are the DNA sequences difference among the same species in the level of nucleic acids and are widely applied in clinical fields such as personalized medicine. The routine and labor-intensive methods to determine SNPs are performing the sequence homology search by using BLAST and navigating the trace of chromatogram files generated by high-throughput DNA sequencing machine by using Chromas program. In this paper, we developed SNPchaser, a web-based program for detecting SNPs substitution and heterozygosity existence, to improve the labor-intensive method in determining SNPs. SNPchaser performed sequence alignment and visualized the suspected region of SNPs by using user's reference sequence, AB1 files, and positional information of SNPs. It simultaneously provided the results of sequences alignment and chromatogram of relevant area of SNPs to user. In addition, SNPchaser can easily determine existence of heterozygosity in SNPs area. SNPchaser is freely accessible via the web site http://www.bioinformatics.ac.kr/SNPchaser and the source codes are available for academic research purpose.

Cloning and Expression of a Serine Proteinase Gene Fragment from Acanthamoeba culbertsoni

  • Park, Ki-Won;Kim, Tong-Soo;Na, Byoung-Kuk;Song, Chul-Yong
    • BMB Reports
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    • v.31 no.3
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    • pp.303-306
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    • 1998
  • Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.

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cDNA Sequence and mRNA Expression of a Putative Glutathione S-Transferase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Jin-Won;Ryu, Kang-Sun;Ahn, Mi-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.2
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    • pp.157-162
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    • 2003
  • The glutathione S-transferase (GSTs) are enzymes responsible for the protection of cells from chemical toxicants and oxidative stress. We describe here the cDNA sequence and mRNA expression of a putative GST from the mole cricket, Gryllotalpa orientalis. The G. orientalis GST cDNA sequences comprised of 621 bp encoding 207 amino acid residues. The multiple sequence alignment of G. orientalis GST gene with other known insect GSTs showed several conserved residues that may be essential for the enzymatic activity of the protein. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis GST gene with other insect GST sequences revealed that the G. orientalis GST gene belongs to class I GST, forming a strong monophyletic group (100% bootstrap value) exclusively for class I GSTs from a diverse insect species. Northern blot analysis confirmed midgut-specific expression at transcriptional level, evidencing the midgut as a site for GST synthesis.

PCR-based markers developed by comparison of complete chloroplast genome sequences discriminate Solanum chacoense from other Solanum species

  • Kim, Soojung;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.79-87
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    • 2019
  • One of wild diploid Solanum species, Solanum chacoense, is one of the excellent resources for potato breeding because it is resistant to several important pathogens, but the species is not sexually compatible with potato (S. tuberosum) causing the limitation of sexual hybridization between S. tuberosum and S. chacoense. Therefore, diverse traits regarding resistance from the species can be introgressed into potato via somatic hybridization. After cell fusion, the identification of fusion products is crucial with molecular markers. In this study, S. chacoense specific markers were developed by comparing the chloroplast genome (cpDNA) sequence of S. chacoense obtained by NGS (next-generation sequencing) technology with those of five other Solanum species. A full length of the cpDNA sequence is 155,532 bp and its structure is similar to other Solanum species. Phylogenetic analysis resulted that S. chacoense is most closely located with S. commersonii. Sequence alignment with cpDNA sequences of six other Solanum species identified two InDels and 37 SNPs specific sequences in S. chacoense. Based on these InDels and SNPs regions, four markers for distingushing S. chacoense from other Solanum species were developed. These results obtained in this research could help breeders select breeding lines and facilitate breeding using S. chacoense in potato breeding.

A Study on Clustering and Identifying Gene Sequences using Suffix Tree Clustering Method and BLAST (서픽스트리 클러스터링 방법과 블라스트를 통합한 유전자 서열의 클러스터링과 기능검색에 관한 연구)

  • Han, Sang-Il;Lee, Sung-Gun;Kim, Kyung-Hoon;Lee, Ju-Yeong;Kim, Young-Han;Hwang, Kyu-Suk
    • Journal of Institute of Control, Robotics and Systems
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    • v.11 no.10
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    • pp.851-856
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    • 2005
  • The DNA and protein data of diverse species have been daily discovered and deposited in the public archives according to each established format. Database systems in the public archives provide not only an easy-to-use, flexible interface to the public, but also in silico analysis tools of unidentified sequence data. Of such in silico analysis tools, multiple sequence alignment [1] methods relying on pairwise alignment and Smith-Waterman algorithm [2] enable us to identify unknown DNA, protein sequences or phylogenetic relation among several species. However, in the existing multiple alignment method as the number of sequences increases, the runtime increases exponentially. In order to remedy this problem, we adopted a parallel processing suffix tree algorithm that is able to search for common subsequences at one time without pairwise alignment. Also, the cross-matching subsequences triggering inexact-matching among the searched common subsequences might be produced. So, the cross-matching masking process was suggested in this paper. To identify the function of the clusters generated by suffix tree clustering, BLAST was combined with a clustering tool. Our clustering and annotating tool is summarized as the following steps: (1) construction of suffix tree; (2) masking of cross-matching pairs; (3) clustering of gene sequences and (4) annotating gene clusters by BLAST search. The system was successfully evaluated with 22 gene sequences in the pyrubate pathway of bacteria, clustering 7 clusters and finding out representative common subsequences of each cluster

An effcient algorithm for multiple sequence alignment (복수 염기서열 정렬을 위한 한 유용성 알고리즘)

  • Kim, Jin;Song, Min-Dong
    • Proceedings of the Korean Information Science Society Conference
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    • 1998.10c
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    • pp.51-53
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    • 1998
  • 3개 이상의 DNA 혹은 단백질의 염기서열을 정렬하는 복수 염기서열 정렬(multiple sequence alignment)방법은 염기서열들 사이의 진화관계, gene regulation, 단백질의 구조와 기능에 관한 연구에 필수적인 도구이다. 복수 염기서열 정렬문제는 NP-complete 문제군에 속하며, 이 문제를 해결하기 위하여 가장 유용하게 사용되는 알고리즘으로는 dynamic programming이 있다. Dynamic programming은 주어진 입력 염기서열 군들에 대한 최적의 정렬을 생산할 수 있다. 그러나 dynamic programming의 단점은 오랜 실행시간이 요구되며, 때로는 dynamic programming의 속성 때문에 이 알고리즘을 사용하여도 주어진 입력 염기서열 군들에 대한 최적의 정렬을 얻어내지 못하는 경우가 있다. 본 연구에서는 이러한 dynamic programming의 문제를 해결하기 위하여 genetic algorithm을 복수 염기서열 정렬문제에 적용하였다. 본 논문에서는 genetic algorithm의 design과 적용방법을 기술하였다. 본 연구에서 제안된 genetic algorithm을 사용하여 dynamic programming의 단점이었던 오랜 실행시간을 줄일 수 있었으며, dynamic programming이 제공하지 못하는 최적의 염기서열 정렬을 제공할 수 있었다.

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Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

cDNA Sequence and mRNA Expression of a Putative Alcohol Dehydrogenase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Lee, Young-Sin;Ryu, Kang-Sun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.1
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    • pp.37-44
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    • 2003
  • Alcohol dehydrogenases (AHDs) are enzymes responsible for the catalysis of the reversible conversion of various alcohols to their corresponding aldehydes and ketonesis. Until now cDNA sequences of ADH gene is informed exclusively from several diptean species. We describe here the cDNA sequence and mRNA expression of a putative ADH gene from the mole cricket, Gryllotalpa orientalis, and phylogenetic relationships among known insect ADHs. The G. orientalis ADH cDNA sequences comprised of 798 bp encoding 266 amino acid residues. The multiple sequence alignment of G. orientalis ADH gene and known dipteran ADHs shared 100% identity in the nine amino acid residues that are important for the enzymatic activity in Drosophila melanogaster. Percent sequence identity ranged from 25% to 32% among all insect ADHs including both types of ADHs. G. orientalis ADH gene showed no clear resemblance to any dipteran species and type. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis ADH gene with available dipteran ADH genes including both types of ADHs further confirmed that the G. orientalis ADH gene is not clearly assigned to either type of ADHs. Northern blot analysis revealed a stronger signal in the fat body than midgut and epidermis, indicating that the fat body possibly is a main site for the synthesis of the G. orientalis ADH protein.