• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.79-84
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• 2002

Suppression subtractive hybridization (SSH) was used to isolate wound-induced cDNAs from wounded soybean. One of wound-induced cDNA, gmwi33 showed high homology with genes encoding $\beta$-amyrin synthase. The full length cDNA of gmwi33, designated GmAMS1, is 2416 bp long and contains an open reading frame consisted of 739 amino acids. GmAMS1 protein showed 89% identity with licorice GgbAS1 and 86% identity with pea OSCPSY. In 5 day-old, dark-grown seedlings, the expression of GmAMS1 was most strongly induced by light and weakly induced by methyl jasmonate and by low temperature. However, GmAMS1 was not induced by elicitor or UV-B treatment. Such expression pattern might be closely related with the oxygen-radical scavenging activity of soyasaponin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.3
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• pp.354-361
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• 2015

Mitochondrial heat shock protein 75 (mtHSP75) is a member of the HSP90 family and plays essential roles in refolding proteins of the mitochondrial matrix. Mitochondria provide energy in the form of ATP and generate reactive oxygen species (ROS). Heat shock proteins (HSPs) are activated in response to stress, and protect cells. In this study, we characterized the mtHSP75 of the big-belly seahorse Hippocampus abdominalis. The protein (BsmtHSP75) is encoded by an open reading frame (ORF) of 2,157 nucleotides, has 719 amino acids (aa), and is of molecular mass 82 kDa. BsmtHSP75 has two functional domains, a histidine kinase-like ATPase (HATPase_c) domain (123-276 aa) and an HSP90 family domain (302-718 aa). BsmtHSP75 was expressed in all tested tissues of healthy seahorses. The ovary contained the highest transcription level, followed (in order) by the blood, brain, and muscle. Pouch tissue showed the lowest expression level. The expression of BsmtHSP75 was significantly (P<0.05) up-regulated on viral or bacterial challenge, suggesting that BsmtHSP75 plays a role in the immune defense against bacterial and viral pathogens.

• Korean Journal of Microbiology
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• v.54 no.2
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• pp.152-154
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• 2018

Klebsiella pneumoniae is a Gram-negative, rod-shape bacterium causing disease in human and animal lungs. K. pneumoniae has been often found to gain antimicrobial resistance, thus it has been difficult to treat K. pneumoniae infection with antibiotics. For such infection, bacteriophage can provide an alternative approach for pathogenic bacterial infection with antimicrobial resistance, because of its sensitivity and specificity to the host bacteria. Bacteriophage KP1 was isolated in sewage and showed specific infectivity to K. pneumoniae. Here, we report the draft genome sequence of Klebsiella pneumoniae phage KP1. The draft genome of KP1 is 167,989 bp long, and the G + C content is 39.6%. The genome has 295 predicted ORFs and 14 tRNA genes. In addition, it encodes various enzymes which involve in lysis of the host cell such as lysozyme and holin.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.185-197
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• 2003

This study was performed to identify the characteristics of the OFC1 gene (locus: chromosome 6p24.3) in Korean patients, which is assumed to be the major gene behind the nonsyndromic cleft lip and palate. The sample consisted of 80 subjects: 40 nonsyndromic cleft lip and palate patients (proband, 20 males and females, mean age 14.2 years); and 40 normal adults (20 males and 20 females, mean age 25.6 years). Using PCR-based assay, the OFC1 gene was amplified, sequenced, and then searched for similar protein structures. Results were as follows: 1. The OFC1 gene contains the microsatellite marker 'CA' repeats. The number of the reference 'CA' repeats was 21 times, and formed as TA(CA)11TA(CA)10. But, in Koreans, the number of tandem 'CA' repeats was varied from 17 to 26 except 18, and 'CA' repeats consisted of TA(CA)n. 2. Nine allelic variants were found. Distribution of the OFC1 allele was similar between the patients and control group. 3. There was a replacement of the base 'T' to 'C' after 11 tandem 'CA' repeats in Koreans compared with Weissenbach's report. However, the difference did not seem to be the ORF prediction results between Koreans and Weissenbach's report. 4. The BLAST search results showed the Telomerase reverse transcriptase (TERT) and the Nucleotide binding protein 2 (NBP2) as similar proteins. The TERT was a protein product by the hTERT gene in the locus 5p15.33 (NCBI Genome Annotation; NT023089) The NBP2 was a protein product by the ABCC3 (ATP-binding cassette, sub-family C) gene in the locus 17q22 (NCBI Genome Annotation; NT010783). 5. In the Pedant-Pro database analysis, the predictable protein structure of the OFC1 gene had at least one transmembrane region and one non-globular region.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.63-70
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• 2018

Brazzein is the smallest sweet protein and was isolated from the fruit pulp of Pentadiplandra brazzeana Baillon, native to tropical Africa. From ancient times, the indigenous people used this fruit in their diet to add sweetness to their daily food. Brazzein is 500 to 2000 times sweeter than sucrose on a weight basis and 9500 times sweeter on a molar basis. This unique property has led to increasing interest in this protein. However, it is expensive and difficult to produce brazzein other than in its native growing conditions which limits its availability for use as a food additive. In this study, we report high production yields of, brazzein protein in transgenic rice plants. An ORF region encoding brazzein and driven by the $2{\times}CaMV\;35S$ promoter was introduced into rice genome (Oryza sativa Japonica) via Agrobacterium-mediated transformation. After transformation, 17 regenerated plant lines were obtained and these transgene-containing plants were confirmed by PCR analysis. In addition, the selected plant lines were analyzed by Taqman PCR and results showed that 9 T0 lines were found to have a single copy out of 17 transgenic plants. Moreover, high and genetically stable expression of brazzein was confirmed by western blot analysis. These results demonstrate that recombinant brazzein was efficiently expressed in transgenic rice plants, and that we have developed a new rice variety with a natural sweetener.

• The Korean Journal of Malacology
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• v.30 no.4
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• pp.399-408
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• 2014

Glutathione S-transferases (GSTs) are a superfamily of detoxification enzymes that primarily catalyze the nucleophilic addition of reduced glutathione to both endogenous and exogenous electrophiles. In this study, we isolated and characterized a full-length of alpha class GST cDNA from the abalone (Haliotis discus hannai). The abalone GST cDNA encodes a 223-amino acid polypeptide with a calculated molecular mass of 25.8 kDa and isoelectric point of 5.69. Multiple alignments and phylogenetic analysis with the deduced abalone GST protein revealed that it belongs to the alpha class GSTs and showed strong homology with disk abalone (Haliotis discus discus) putative alpha class GST. Abalone GST mRNA was ubiquitously detected in all tested tissues. GST mRNA expression was comparatively high in the mantle, gill, liver, and digestive duct, however, lowest in the hemocytes. Expression level of abalone GST mRNA in the mantle, gill, liver, and digestive duct was 182.7-fold, 114.8-fold, 4675.8-fold, 406.1-fold higher than in the hemocytes, respectively. Expression level of abalone GST mRNA in the liver was peaked at 6 h post-infection with Vibrio parahemolyticus and decreased at 12 h post-infection. While the expression level of abalone GST mRNA in the hemocytes was drastically increased at 3 h post-infection with Vibrio parahemolyticus. These results suggest that abalone GST is conserved through evolution and may play roles similar to its mammalian counterparts.

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.39-44
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• 2014

We isolated highly-expressed genes in the posterior silk glands of silkworm on a previously study, which one of these was identified as RNA binding protein-1 homologue (RBP-1) gene. In this study, we investigated gene expressional characteristics of the RBP-1 depending on silkworm development stages and several tissues of the larvae, respectively. Northern blot hybridization analysis showed that the RBP-1 gene was expressed high in larval and pupal periods, and highly expressed than endogenous internal control gene (BmA3) on all tested larval tissues. In addition, we isolated and analyzed a phage DNA having 1,660 bp-long promoter region of the RBP-1 gene from a genomic DNA library. To study the RBP-1 gene promoter activity, RBP-1 (-740/+ 30) was amplified by PCR and subcloned into a pGL3 basic vector to generate pGL-RBP1. A luciferase report vector carrying RBP-1 gene promoter (770 bp) was tested by luciferase assay in Sf9 cells. In the result, the RBP-1 gene promoter was more efficient than constitutive promoter (BmA3) by approximately ten percent.

• Research in Plant Disease
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• v.22 no.4
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• pp.289-292
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• 2016

We surveyed the occurrence of Japanese yam mosaic virus (JYMV) on Yam in Gyeongsangbukdo pronvince from 2013 to 2015. The symptoms of JYMV were yellow stripes and chlorosis in yam leaves and the infection rate was ranged from 33.6% to 40.8%. We determined nucleotide sequence encoding the polyprotein of JYMV isolate BRI from yam leaves using next-generation sequencing (NGS) method. The partial nucleotide portion (7,736 nucleotides) of the genomic RNA of the JYMV isolate BRI has been sequenced (accession No. KU309315). The region sequenced includes a single open reading frame (ORF) encoding a polyprotein composed of 2,497 amino acids containing the coat protein (CP) and 3' untranslated region (UTR). The genomic organization of this isolate shows almost the same to that of other members of JYMV. The JYMV isolate BRI showed 77% to 79% nucleotide identity with the Japanese and Chinese strains and isolates. This is the first report of the genome nucleotide sequence of JYMV from Dioscorea opposita in Korea.

• Journal of Life Science
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• v.20 no.8
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• pp.1193-1200
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• 2010

The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.