• Title/Summary/Keyword: Protein sequence search

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Functional Analysis of ESTs from the Flower Bud of Korean Ginseng

  • Yang, Deok-Chun;In, Jun-Gyo;Kim, Moo-Sung;Jeon, Jong-Seong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.124-124
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    • 2003
  • In order to study gene expression in a reproductive organ, we constructed a cDNA library of immature flower buds in Korean ginseng and generated expressed sequence tags (ESTs) of 3,360 clones randomly selected. The ESTs could be clustered into 1,844 non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 1,254 groups show similarity to genes of known function. These ESTs clones were divided into sixteen categories depending upon gene function. The most abundant transcripts were unknown protein (72), chlorophyll a/b-binding protein (48), and stylar glycoprotein. There are no useful informations of gene expression during the development of flower bud in Korean ginseng. These results could help to understand the development of flower bud in Korean ginseng.

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Molecular Characterization of Plasmid from Bifidobacterium longum

  • Park, Myeong-Soo;Moon, Hye-Won;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.457-462
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    • 2003
  • The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.

Molecular Cloning and Characterization of the Gene encoding $\beta$-N-acetylhlucosaminidases Homologue from Bombyx mandarina (멧누에(Bombyx mandarina)로부터 $\beta$-N-Acetyglunosamicidase를 코딩하는 cDNA의 분리 및 염기서열 결정)

  • 구태원;황재삼;성규병;윤은영;방혜선;권오유
    • Journal of Sericultural and Entomological Science
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    • v.41 no.3
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    • pp.147-153
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    • 1999
  • Chitinolytic enzymes such as ${\beta}$-N-acetylglucosaminidase are major hydrolases involved in insect molting. We have isolated, sequenced a CDNA encoding ${\beta}$-N-acetylglucosaminidase from the silkworm, Bombyx mandarina, and compared its sequence with genes encoding chitinolytic enyzmes from other sources. The insert DNA in the clone is 3,284 nucleotides long with an open reading frame of 1,788 nucleotides that encodes a protein of 596 amino acids with a molecular weight of 68.2 kDa. There is a 3’-untranslated region composed with 1.479 nucleotides and are several potential polyadenylation signals. The predicted amino acid sequence apparently contains a leader peptide of 23 amino acids. A search of the amino acids sequence databases for sequences similarities to other ${\beta}$-N-acetylglucosaminidases or ${\beta}$-N-acetylhexosaminidases. The highest similarity matched with the enzyme from B. mori, which has a sequence identity of 95%. On the other hand, the identity between the B. mandarina enzyme and those from M. sexta and human are 70% and 24%, respectively.

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Isolation and Characterization of Pathogenesis-Related Protein 5 (PgPR5) Gene from Panax ginseng

  • Kim, Yu-Jin;Lee, Jung-Hye;Jung, Dae-Young;Sathiyaraj, Gayathri;Shim, Ju-Sun;In, Jun-Gyo;Yang, Deok-Chun
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.400-407
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    • 2009
  • A pathogenesis-related protein (PgPR5) gene that isolated from the leaf of Panax ginseng was characterized. The ORF is 756 bp with a deduced amino acid sequence of 251 residues. The calculated molecular mass of the matured protein is approximately 27.5 kDa with a predicated isoelectric point of 7.80. A GenBank BlastX search revealed that the deduced amino acid of PgPR5 shares highest sequence similarity to PR5 of Actinidia deliciosa (80% identity, 87% similarity). PgPR5 has a C-terminal and N-terminal signal peptide, suggesting that it is a vacuolar secreted protein. The expression of PgPR5 under various environmental stresses was analyzed at different time points using real-time PCR. Our results reveal that PgPR5 is induced by salt stress, chilling stress, heavy metal, UV, and pathogen infection. These results suggest that the PgPR5 could play a role in the molecular defence response of ginseng to abiotic and pathogen attack. This is the first report of the isolation of PR5 gene from the P. ginseng.

Characterization of the xaiF Gene Encoding a Novel Xylanase-activity- increasing Factor, XaiF

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.378-387
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    • 1998
  • The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 [about l-kb region downstream from the translational termination codon (TAA) of the xynA gene]was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

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HCoV-IMDB: Database for the Analysis of Interactions between HCoV and Host Immune Proteins

  • Kim, Mi-Ran;Lee, Ji-Hae;Son, Hyeon Seok;Kim, Hayeon
    • International journal of advanced smart convergence
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    • v.8 no.1
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    • pp.1-8
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    • 2019
  • Coronaviruses are known respiratory pathogens. In the past, most human coronaviruses were thought to cause mild symptoms such as cold. However recently, as seen in the Severe Acute Respiratory Syndrome (SARS) and the Middle East Respiratory Syndrome (MERS), infectious diseases with severe pulmonary disease and respiratory symptoms are caused by coronaviruses, making research on coronaviruses become important. Considering previous studies, we constructed 'HCoV-IMDB (Human Corona Virus Immune Database)' to systematically provide genetic information on human coronavirus and host immune information, which can be used to analyze the interaction between human coronavirus and host immune proteins. The 'HCoV-IMDB' constructed in the study can be used to search for genetic information on human coronavirus and host immune protein and to download data. A BLAST search specific to the human coronavirus, one of the database functions, can be used to infer genetic information and evolutionary relationship about the query sequence.

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

Expression Analysis of ESTs Derived from the Four-Year Root of Chunpoong (Panax ginseng C.A. Meyer)

  • Yang, Deok-Chun;In, Jun-Gyo;Lee, Bum-Soo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.121-121
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    • 2003
  • Expressed sequence tags (EST) are help to quickly identify functions of expressed genes and to understand the complexity of gene expression. To assist genetic study of the root development in Panax ginseng, which is one of the most important medicinal plant, expressed sequence tags (EST) analysis was carried out. We constructed a CDNA library using the 4-year Chunpoon root. Partial sequences were obtained from 3,841 clone. The ESTs could be clustered into 2,056 (64%) non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 1,498 groups show similarity to genes of known function. These ESTs clones were divided into eighteen categories depending upon gene function. The most abundant transcripts were major latex protein (41), ribonuclease 2 (36), metallothionein 2(35). Our extensive EST analysis of genes expressed in 4-year Chunpoong root not only contributes to the understanding of the dynamics of genome expression patterns in root organ development but also adds data to the repertoire of all genomic genes.

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MODf : An Effective and Fast Algorithm for Identification of PTM in Large Protein Sequence Database (MODf : 대규모 단백질 DB에서 효과적이고 빠르게 PTM을 동정하는 알고리즘)

  • Shin, Seong-Ho;Park, Heejin;Paek, Eunok
    • Annual Conference of KIPS
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    • 2010.11a
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    • pp.1834-1836
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    • 2010
  • 프로테오믹스는 세포 안 또는 개체 안의 모든 단백질을 총체적으로 연구하는 분야이다. 단백질 동정은 단백질이 어떤 아미노산의 서열로 구성되었는지를 확인하는 것이다. 하지만 Post-translational modification과 같은 단백질 변형을 고려하게 되면 단백질 동정은 매우 어렵게 된다. $MOD^i$ 알고리즘은 단백질 동정을 할 때 Post-translational modification의 종류나 개수에 제한 없이 단백질 동정을 정확하게 수행한다. 하지만, 대용량 단백질 서열 데이터베이스를 사용하면 수행시간이 많이 걸리는 단점이 있다. 본 논문에서는 $MOD^i$를 보완하기 위해 대용량 데이터베이스에서 후보 단백질을 선정하는 알고리즘을 통해서 개선된 $MOD^f$ 알고리즘을 제안하고 Target-decoy search strategy를 적용하여 정확성을 분석한다. 후보 단백질 선정 알고리즘과 Target-decoy search strategy 적용 결과 $MOD^f$$MOD^i$에 비해 정확도를 희생하지 않으면서 수행속도는 약 2배 향상되었다.

Sequence-Based Screening for a Putative ${\gamma}$-Butyrobetaine Hydroxylase Gene from Neurospora crassa

  • Hur Min-Sang;Cho Jae-Yong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1468-1471
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    • 2006
  • The last step in L-carnitine biosynthesis in eukaryotic organisms is mediated by ${\gamma}$-butyrobetaine hydroxylase (EC1.14.11.1), a dioxygenase that converts ${\gamma}$-butyrobetaine to L-carnitine. This enzyme was previously identified from rat liver and humans, and the peptide sequence of human ${\gamma}$-butyrobetaine hydroxylase was used to search the Neurospora crassa genome database, which led to an identification of an open reading frame (ORF) consisting of 1,407 bp encoding a polypeptide of 468 amino acids. When this protein was expressed in Saccharomyces cerevisiae, the crude cell-free extract exhibited ${\gamma}$-butyrobetaine hydroxylase activity.