• BMB Reports
• /
• 제34권4호
• /
• pp.371-378
• /
• 2001

The Glycoprotein D (gD) gene of the HSV-1 strain F was cloned, sequenced, recombinated into the HcNPV (Hyphantria cunea nuclear polyhedrosis virus) expression vector and expressed in insect cells. The gD gene was located in the 6.43 kb BamHI fragment of the strainF. The open reading frame (ORF) of the gD gene was 1,185 by and codes 394 amino acid residues. Recombinant baculoviruses, GD-HcNPVs, expressing the gD protein were constructed. Spodoptera frugiperda cells, infected with the recombinant virus, synthesized a matured gX-gD fusion protein with an approximate molecular weight of 54 kDa and secreted the gD proteins into the culture media by an immunoprecipitation assay The fusion gD protein was localized on the membrane of the insect cells, seen by using an immunofluorescence assay The deduced amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. These results indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins.

• The Plant Pathology Journal
• /
• 제23권4호
• /
• pp.245-250
• /
• 2007

The complete genome sequence of Zucchini yellow mosaic virus stain A (ZYMV-A) isolated from a hollyhock (Althaea rosea) was determined by using RT-PCR with a series of primer sets. The virus genome consisted of 9593 nucleotides (nt), excluding the poly(A) tract at 3' terminus of the virus genome, with 5' and 3' untranslated region of 139 and 211 nt, respectively. The deduced polyprotein of ZYMV-A consisted of 3080 amino acid (aa) residues and was 351 kDa in molecular weight. All proteolytic cleavage sites of the polyprotein of ZYMV-A were compared with those of ZYMV strains, which showed the cleavage sites were conserved among ZYMV strains. The HC-Pro contained the KITC and PTK motifs, and the DAG motif was located at CP ORF of ZYMV-A, suggesting that ZYMV-A is aphid-transmissible. Phylogenetic tree analysis based on the complete genome among ZYMV strains or CP ORFs with other potyviruses showed ZYMV strains formed a distinct group. These results clearly confirmed that ZYMV-A was another distinct strain in ZYMV population at molecular level.

• Animal cells and systems
• /
• 제12권3호
• /
• pp.149-155
• /
• 2008

We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

• Journal of Microbiology and Biotechnology
• /
• 제24권11호
• /
• pp.1525-1535
• /
• 2014

The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

• 한국항해항만학회:학술대회논문집
• /
• 한국항해항만학회 2006년도 International Symposium on GPS/GNSS Vol.1
• /
• pp.343-348
• /
• 2006

Number of researches on efficient terminal operation system applying RFID(Radio Frequency Identification) are on progress. However, RFID has limitations on tracking and providing accurate positions of containers. In this paper, to enhance the performance of the integrated terminal operation system, an efficient loading management of yard is proposed by applying RFID-based RTLS(Real Time Locating System) that provides real-time accurate positions of containers. We found that a group-based sequence system is more efficient than the existing individual sequence number system in the container yard. In the group-based sequence system, the containers in the same group should have similar characteristics such as port of destination(POD), size, weight, etc. In order to run this system, we have proposed the parameters to the unspecified N bytes of RFID tag which are specified in ISO 18000-7. And, this paper proposed a development of integrated operation system of container terminal using RFID for reducing the ship turnaround time in ubiquitous port environment.

• Fisheries and Aquatic Sciences
• /
• 제24권4호
• /
• pp.163-170
• /
• 2021

Peptides are naturally occurring biological molecules that are found in all living organisms. These biologically active peptides play a key role in various biological processes. The aim of this study is the extraction and the purification of bioactive materials that induce relaxation of an apical muscle from the pyloric caeca of Patiria pectinifera. The acidified pyloric caeca extract was partially separated by the solid phase extraction using a stepwise gradient on Sep-Pak C18 cartridge. Among the fractions, materials eluted with 60% methanol/0.1% trifluoroacetic acid was put a thorough of a series of high performance liquid chromatography (HPLC) steps to isolate a neuropeptide with relaxation activity. The purified compound was eluted at 28% acetonitrile in 0.1% trifluoroacetic acid with retention time of 25.8 min on the CAPCELL-PAK C18 reversed-phase column. To determine the molecular weight and the amino acid sequence of the purified peptide, LC-MS and Edman degradation method were used, respectively. The primary structure of the peptide was determined to be FGMGGAYDPLSAGFTD which corresponded to the amino acid sequence of a starfish myorelaxant peptide (SMP) isotype (SMPb) found in the cDNA sequence encoding SMPa and its isotypes. In this study, a muscle relaxant neuropeptide (SMPb) has been isolated from pyloric caeca of starfish P. pectinifera. This is the first report of SMPb isolation on the protein level from P. pectinifera.

• 한국해양생명과학회지
• /
• 제3권1호
• /
• pp.1-8
• /
• 2018

Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA²⁺ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

• 한국소성가공학회:학술대회논문집
• /
• 한국소성가공학회 2007년도 춘계학술대회 논문집
• /
• pp.397-400
• /
• 2007

Recently, rheology forming technology has been interested in industrial and academic for light weight materials and to solve environmental issues. In this study, the rheology material production equipments were used to be made spiral shape by special design. And the experiment variables established stirring time 0 $\sim$ 1200 sec, stirring velocity 0 $\sim$ 100 rpm and several material temperature of semi - solid states. The rheology materials were made for established experiment conditions then measured mechanical properties. Sequence-production equipments were appended to fabrication system of rheology material for make rheology materials continually. Therefore, the development of sequence-production equipments were demanded for fine grains and for uniform globule shape rheology materials by a specially designed spiral stirrer machine.

• 한국통신학회논문지
• /
• 제38A권5호
• /
• pp.416-425
• /
• 2013

A hybrid PN code phase acquisition system based on a least-mean-square adaptive filter, interpreted as a channel estimator is proposed and analyzed for direct-sequence spread-spectrum systems under frequency selective Rayleigh fading channels. Closed form expressions are derived for the filter tap weights and detection/false alarm probabilities. Compared to previously proposed systems, the proposed system achieves smaller mean acquisition times, is more robust to the operating signal-to-noise ratio and allows for multiplication free tap weight updates.

• Animal cells and systems
• /
• 제13권3호
• /
• pp.345-350
• /
• 2009

We have isolated and characterized a new insect chicken type (c-type) lysozyme gene from the common cutworm, Spodoptera litura. The full-length cDNA of Spodoptera lysozyme is cloned by rapid amplification of cDNA ends PCR (RACE-PCR). The isolated cDNA consists of 1039 bp including the coding region for a 142-amino acid residue polypeptide, which included a signal peptide of 21-amino acid residue and a mature protein of 121-amino acid residue. The predicted molecular weight of mature lysozyme and its theoretical isoelectric point from amino acid composition is 13964.8 Da and 9.05, respectively. The deduced amino acid sequence of Spodoptera lysozyme gene shows the highest similarity (96.7%) to Spodoptera exigua lysozyme among other lepidopteran species. Amino acid sequence comparison with other the c-type lysozymes, Spodoptera lysozyme has the completely conserved $Glu^{32}$ and $Asp^{50}$ of the active site and eight Cys residues are completely conserved in the same position as that of other lepidopteran lysozymes.