• The KIPS Transactions:PartD
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• v.10D no.2
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• pp.261-266
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• 2003

A protein, which is a linear polymer of amino acids, is one of the most important bio-molecules composing biological structures and regulating bio-chemical reactions. Since the characteristics and functions of proteins are determined by their amino acid sequences in principle, protein sequence determination is the starting point of protein function study. This paper proposes a protein sequence prediction method based on data mining techniques, which can overcome the limitation of previous bio-chemical sequencing methods. After applying multiple proteases to acquire overlapped protein fragments, we can identify candidate fragment sequences by comparing fragment mass values with peptide databases. We propose a method to construct multi-partite graph and search maximal paths to determine the protein sequence by assembling proper candidate sequences. In addition, experimental results based on the SWISS-PROT database showing the validity of the proposed method is presented.

• The Plant Pathology Journal
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• v.17 no.3
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• pp.115-122
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• 2001

Potato virus X (PVX) is a flexuous rod-shaped virus containing a single plus-strand RNA. Viral RNA synthesis is precisely regulated by regulatory viral sequences and by viral and/or host proteins. RNA sequence element as well as stable RNA stem-loop structure in the 5' end of the genome affect accumulation of genomic RNA and subgenomic RNA (sgRNA). The putative sgRNA promoter regions upstream of the PVX triple gene block (TB) and coat protein (CP) gene were critical for both TB and CP sgRNA accumulation. Mutations that disrupted complementarity between a region at the 5' end of the genomic RNA and the sequences located upstream of each sgRNA initiation site is important for PVX RNA accumulation. Compensatory mutations that restore complementarity restored sgRNA accumulation levels. However, the extent of reductions in RNA levels did not directly correlate with the degree of complementarity, suggesting that the sequences of these elements are also important. Gel-retardation assays showed that the 5' end of the positive-strand RNA formed an RNA-protein complex with cellular proteins, suggesting possible involvement of cellular proteins for PVX replication. Future studies on cellular protein binding to the PVX RNA and their role in virus replication will bring a fresh understanding of PVX RNA replication.

• Journal of fish pathology
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• v.24 no.3
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• pp.297-305
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• 2011

A trap identification system for isolating functional sequences to allow the constitutive expression of foreign protein from Edwardsiella tarda was developed. Using the green fluorescent protein (GFP) reporter-based trap system, various functional sequences to drive heterologous expression of the GFP were selectable in Escherichia coli host. However from the bioinformatic sequence analysis, all the segments predicted as regulatory regions were not native promoters actually existing upstream of endogenous E. tarda genes. Instead, a number of non-authentic sequences, possibly resulted from the random shuffling and/or intermolecular ligation were also proven to be able to display a potent GFP expression in the recombinant E. coli. Further analysis with selected clones showed that both authentic and non-authentic sequences could function in as a constitutive promoter, leading quite a consistent and stable GFP expression after repetitive subcultures. Microscopic examination also confirmed the uniform pattern of GFP expression in every host bacterium. Semi-quantitative assay of GFP showed that there was no clear relationship between expression levels and organizational features of the promoters trapped. Functional promoter-like elements achieved in the present study could be a good starting material for multivalent genetic engineering of E. tarda in order to produce recombinant vaccines in a cost-effective fashion.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.87-92
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• 2003

We describe here the cloning, expression and characterization of a cDNA encoding a putative chemosensory protein (CSP) from the mole cricket, Gryllotalpa orientalis. The G. orientalis chemosensory protein cDNA sequences comprised of 384 bp with 128 amino acid residues. The G. orientalis chemosensory protein showed 75.4% protein sequence identity to the Locusta migratoria CSP, Northern blot analysis revealed that signal was stronger in head than leg and cuticle, indicating that the head part containing antennae is a main site for G. orientalis chemosensory protein synthesis. The cDNA encoding G. orientalis chemosensory protein was expressed as approximately 12 kDa polypeptide in baculovirus-infected insect cells.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.620-627
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• 2004

On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI- TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (enol). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic $\lambda$ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno 1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation.

• Biomolecules & Therapeutics
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• v.8 no.3
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• pp.276-280
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• 2000

In order to identify the domains of the G$_{\alpha}$16 subunit G protein that are responsible for its activation by the Interleukin-8 receptor, a serious of chimeras between G$_{\alpha}$16 and G$_{\alpha}$11 were assessed for their abilities to be activated by these receptors. Co-expression of IL-8 receptor and chimeras in which the carboxyl-terminal regions of G$_{\alpha}$11 were replaced from 30 up to 156 amino acid residues with the corresponding regions of G$_{\alpha}$16 demonstrated that C-terminal 156 amino acid residues of the G$_{\alpha}$16 were not sufficient to confer IL-8 receptor interaction specificity. Testing of a reciprocal serious of chimeras composed of G$_{\alpha}$16 sequences at the amino terminus and G$_{\alpha}$11 sequences at the carboxyl terminals revealed that sequences extending from the amino tar- minus to amino acid 209 of G$_{\alpha}$16 were sufficient to 7ndow the chimera with 75-80% of interaction specificity for 7-8-induced activation. These results suggest th,.7t combined interactions of the C-terminal 30 amino acid residues and certain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.tain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2005.09a
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• pp.156-160
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• 2005

We introduce a fully automatic clustering method to classier candidate paralog clusters from a set of protein sequences within one genome. A set of protein sequences is represented as a set of nodes, each represented by the amino acid sequence for a protein with the sequence similarities among them constituting a set of edges in a graph of protein relationships. We use graph-based clustering methods to identify structurally consistent sets of nodes which are strongly connected with each other. Our results are consistent with those from current leading systems such as COG/KOG and KEGG based on manual curation. All the results are viewable at http://www.cs.rutgers.edu/${\sim}$seabee.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2004.11a
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• pp.101-106
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• 2004

Subcellular localization is a key functional char acteristic of proteins. With the number of sequences entering databanks rapidly increasing, the importance of developing a powerful tool to identify protein subcellular location has become self-evident. In this paper, we introduce a novel method for predic ting protein subcellular locations from protein sequences. The main idea was motivated from the observation that amino acid pair composition data is redundant. By classifying from multiple feature subsets and using many kinds of amino acid pair composition s, we forced the classifiers to make uncorrelated errors. Therefore when we combined the predictors using a voting scheme, the prediction accuracy c ould be improved. Experiment was conducted on several data sets and significant improvement has been achieve d in a jackknife test.

• The Microorganisms and Industry
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• v.12 no.1
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• pp.9-14
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• 1986

We have adopted a genetic approach to identifying those residues and local sequences in a polypeptide chain which play an important role on the folding pathway. Our approach has been to isolate and characterize mutants which specifically alter the folding and subunit association pathway of a polypeptide chain, without altering the native protein. Such mutants distinguish residues involved in the kinetic control of conformation from residues involved in the stability and activity of the native protein. This approach is complementary to the efforts to characterize mutations which alter the stability of the mature protein(6,7,8). It is likely that many residues will have roles in both aspects of the functioning of the polypeptide chain. We thought it likely, however, that at least with large proteins, these aspects might be segregated in different local sequences.

• Journal of Forest and Environmental Science
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• v.28 no.4
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• pp.278-281
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• 2012

Comparative sequence analyses were conducted on complete mtDNA sequences from four small mammal species in Korea and revealed the presence of 30 well conserved sequences in various regions of the complete mtDNA sequences. The conserved sequences were found in 9 regions in protein coding genes, 10 regions in tRNA genes, 10 in rRNA genes, one region in replication origin and 2 regions in D loop. They could be used to design primers for amplifying complete mtDNA sequences of small mammals.