• Journal of Life Science
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• v.12 no.2
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• pp.188-199
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• 2002

A gene coding for xylanase from thermo-tolerant Bacillus pumilus TX703 was cloned into Escherichia coli DH5 $\alpha$ using pUC19. Among 7,400 transformants, four transformants showed clear zones on the detection agar plates containing oat-spells xylan. One of them which showed highest xylanase activity was selected and its recombinant plasmid, named pXES106, was found to carry 2.24 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynK) was determined, xynK gene was found to consist of 1,227 base-pair open reading frame coding for a polypeptide of 409 amino acids with a deduced molecular weight of 48 kDa. The coding sequence was preceded by a putative ribosome binding site, the transcription initiation signals, and cia-acting catabolite responsive element. The deduced amino acids sequence of xylanase is similar to those of the xylanases from Hordeum vulgare (barley) and Clostridium thermocellum, with 39 and 31% identical residues, respectively. The amino acids sequence of this xylanase was quite different from those of the xylanases from other Bacillus species.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.1
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• pp.51-57
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• 2003

A cDNA encoding a putative alcohol dehydrogenase (ADH) class III was cloned from the silkworm, Bombyx mono The full length cDNA is 1,385 nucleotides long and contains an open reading frame of 1,128 bp encoding 376 amino acid residues. The B. mon ADH III protein sequence was aligned with ADH III known from various organisms. Interestingly, the protein sequence of B. mon ADH III showed 87% and 85% identity to ADH III from marine fish Sparus aurata and Branchiostoma floridae, respectively, whereas rather low sequence identity (83%) to Drosophila melanogaster ADH III was observed. Northern blot analysis revealed that B. mon ADH III mRNA is expressed in all tissues from larva examined: fat body, midgut, epidermis, silk gland and ovary, with the highest level found in the fat body.

• BMB Reports
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• v.45 no.1
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• pp.14-19
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• 2012

A feruloyl esterase encoding gene (designated fae6), derived from a leachate metagenomic library, was cloned and the nucleotide sequence of the insert DNA determined. Translational analysis revealed that fae6 consists of a 515 amino acid poly-peptide, encoding a 55 kDa pre-protein. The Fae6 primary structure contained the G-E-S-A-G sequence, which corresponds well with a typical catalytic serine sequence motif (G-x-S-x-G). The fae6 gene was successfully over-expressed in E. coli and the recombinant protein was purified to 8.4 fold enrichment with 17% recovery. The $K_M$ data showed Fae6 has a high affinity to methyl sinapate while thermostability data indicated that fae6 was thermolabile with a half life ($T_{1/2}$) < 30 min at $50^{\circ}C$. High affinity for Fae6 against methyl sinapate, methyl ferulate and ethyl ferulate suggest that the enzyme can be useful in hydrolyzing ferulated polysaccharides in a biorefinery process.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2000.11a
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• pp.45-52
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• 2000

Genomic approach produces massive amount of data within a short time period, New high-throughput automatic sequencers can generate over a million nucleotide sequence information overnight. A typical DNA chip experiment produces tens of thousands expression information, not to mention the tens of megabyte image files, These data must be handled automatically by computer and stored in electronic database, Thus there is a need for systematic approach of data collection, processing, and analysis. DNA sequence information is translated into amino acid sequence and is analyzed for key motif related to its biological and/or biochemical function. Functional genomics will play a significant role in identifying novel drug targets and diagnostic markers for serious diseases. As an enabling technology for functional genomics, bioinformatics is in great need worldwide, In Korea, a new functional genomics project has been recently launched and it focuses on identi☞ing genes associated with cancers prevalent in Korea, namely gastric and hepatic cancers, This involves gene discovery by high throughput sequencing of cancer cDNA libraries, gene expression profiling by DNA microarray and proteomics, and SNP profiling in Korea patient population, Our bioinformatics team will support all these activities by collecting, processing and analyzing these data.

• Journal of fish pathology
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• v.23 no.3
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• pp.429-440
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• 2010

We constructed a rock bream (Oplegnathus fasciatus) gill cDNA library and a total of 1450 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 1450 EST clones, 1022 EST clones showed significant homology to previously described genes while 428 ESTs were unidentified, and 259 clones were hypothetical, or unnamed proteins. Encoding 313 different sequences were identified as putative bio-defense genes or genes associated with immune response.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.360-365
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• 1992

Clinical isolates of 8 ofloxacin resistant Staphylococcus auresu (ORSA) were subjected to MIC test, Southern analysis on gyrA locus and nucleotide sequence analysis of 290 bp of gyrA gene (gyrA-290) spanning amino acid 26 to 121 in order to understand the mechanism of quinolone resistance in Staphylococcus aureus. ORSAs showed highlevel resistance against quinolones (8-250 fold increase of MICs) and also significant resistance agianst ${\beta}-lactams$ (2-32 fold increase of MICs). However, ORSs did not show any change in sensitivity agianst vancomycin. Southern analysis of ORSAs with HindIII, PstI and AluI revealed RFLPs on gyrA locus. In order to further analyze the gyrA gene, gyrA-290 was amplified by PCR and cloned to pTZ vector. Subsequent nucleic acid sequence analysis of gyrA-290 demonstrated a point mutation of C to T resulting amino acid change of Ser-84 to Leu-84 in all 8 ORSA strains. The substitution at 84th amino acid of tyrase A might confer one mechanism of high level quinolone resistance in Staphylococcus aureus.

• BMB Reports
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• v.30 no.3
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• pp.173-176
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• 1997

We isolated a cDNA clone that encodes a ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) from spinach using a soybean rbcS cDNA as a probe. The small subunit consists of 180 amino acids including a transit peptide of 57 residues. Comparison of the amino acid sequence with those of other plant species shows a maximum of 70-80% identical residues. Southern blot analysis suggests the existence of multiple rbcS genes in the spinach genome. Northern blot analysis indicates that the rbcS gene is expressed predominantly in leaves and that the expression of the gene is induced by light.

• Journal of Plant Biology
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• v.38 no.2
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• pp.137-141
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• 1995

We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.713-714
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• 2000

Six monoclonal antibodies to human cardiac troponin I (hcTnI) were produced to eventually develop an immunosensor for acute myocardial infarction (AMI). For the characterization of these antibodies, a set of 11 different peptides covering selected ranges of the complete amino acid sequence of hcTnI was prepared and used for epitope mapping. Such analysis allowed to select an appropriate pair of antibodies that can form a sandwich type of immune complexes and was consequently used for an immunoassay.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.508-514
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• 2001

A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.