• Korean Journal of Microbiology
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• v.39 no.1
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• pp.1-7
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• 2003

The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

• Journal of Ginseng Research
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• v.32 no.4
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• pp.300-304
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• 2008

Cysteine proteinases play an essential role in plant growth and development but also in senescence and programmed cell death. They participate in both anabolic and catabolic processes. In addition, they are involved in signalling pathways and in the response to biotic and abiotic stresses. A cDNA clone encoding cysteine proteinase (CP) gene, designated PgCysP1, was isolated from Panax ginseng C. A. Meyer. Reverse transcriptase (RT)-PCR results showed that PgCysP1 expressed at different level in P. ginseng hairy root. Different stresses such as biotic as well as abiotic stresses triggered a significant induction of PgCysP1. The positive responses of PgCysP1 to the various stimuli suggested that PgCysP1 may help to protect the plant against reactive environmental stresses.

• Journal of Animal Science and Technology
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• v.50 no.2
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• pp.265-272
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• 2008

The CCAAT/enhancer binding protein β(C/EBPβ), a member of the leucine zipper DNA-binding protein of transcription factor, plays a crucial role in the control of early phases of adipocyte differentiation. In this studies, we report the identification, characterization, and expression of the Korean native cattle C/EBPβ gene. The Korean native cattle and black cattle C/EBPβ cDNA includes a 1047bp open reading frame encoding a protein of 348 amino acids. The C/EBPβ cDNA sequence of the Korean native cattle and black cattle shows high conservation with the corresponding amino acid sequences reported in other species. The distribution of C/EBPβ mRNA in various tissues of Korean native cattle aged 26 months was investigated using Northern Blot analysis. The C/EBPβ expression was detected in adipose tissue, lung, sirloin while expression was not detected in heart, kidney, small intestine, colon, and liver. However, we are analyzed polymorphism of bZIP domain in the C/EBPβ gene. A polymorphism was not identified at this position.

• Journal of Life Science
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• v.25 no.2
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• pp.121-129
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• 2015

Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

• Journal of Life Science
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• v.27 no.9
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• pp.1003-1010
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• 2017

An open reading frame coding for mannanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by polymerase chain reaction amplification, and completely sequenced. This mannanase gene, designated man26AT, consisted of 3,162 nucleotides encoding a polypeptide of 1,053 amino acid residues. Based on the deduced amino acid sequence, Man26AT was identified as a modular enzyme, which included a catalytic domain belonging to the glycosyl hydrolase family 26 and two carbohydrate-binding modules, CBM27 and CBM11. The amino acid sequence of Man26AT was homologous to that of several putative mannanases, with identity of 81% for P. ihumii and identity of less than 57% for other strains of Paenibacillus. A cell-free extract of recombinant E. coli carrying the man26AT gene showed maximal mannanase activity at $55^{\circ}C$ and pH 5.5. The enzyme retained above 80% of maximal activity after preincubation for 1 h at $50^{\circ}C$. Man26AT was comparably active on locust bean gum (LBG), galactomanan, and kojac glucomannan, whereas it did not exhibit activity on carboxymethylcellulose, xylan, or para-nitrophenyl-${\beta}$-mannopyranoside. The common end products liberated from mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, or LBG by Man26AT were mannose, mannobiose, and mannotriose. Mannooligosacchrides larger than mannotriose were found in enzymatic hydrolyzates of LBG and guar gum, respectively. However, Man26AT was unable to hydrolyze mannobiose. Man26AT was intracellularly degraded into at least three active proteins with different molecular masses by zymogram.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.55 no.2
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• pp.151-158
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• 2010

Camelina sativa L., known as popular names "gold-of-pleasure" or "false flax" is an alternative oilseed crop that can be grown under different climatic and soil conditions. Up to date, however, the genomic information of Camelina has not been studied in detail. Therefore, a cDNA library was constructed and characterized from young leaves. The constructed cDNA library incorporated of 1334 cDNA clones and the size of the insertion fragments average was 736 base pair. We generated a total of 1269 high-quality expressed sequence tags (ESTs) sequences. The result of cluster analysis of EST sequences showed that the number of unigene was 851. According to subsequent analysis, the 476 (55.9%) unigenes were highly homologous to known function genes and the other 375 (44.1%) unigenes were unknown. Remaining 63 (7.4%) unigenes had no homology with any other peptide in NCBI database, indicating that these seemed to be novel genes expressed in leaves of Camelina. The database-matched ESTs were further classified into 17 categories according to their functional annotation. The most abundant of categories were "protein with binding function or cofactor requirement (27%)", "metabolism (11%)", "subcellular localization (11%)", "cellular transport, transport facilities and transport routes (7%)", "energy (6%)", "regulation of metabolism and protein function (6%)". Our result in this study provides an overview of mRNA expression profile and a basal genetic information of Camelina as an oilseed crop.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.515-530
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• 1995

Coronaviridae에 속하는 transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)를 specific하게 detection할 수 있는 방법을 개발하고자 본 연구를 수행하였다. 두 바이러스 모두 RNA 바이러스이기 때문에 reverse transcription-polymerase chain reaction(RT-PCR)으로 nucleocapsid(N) protein gene의 cDNA를 증폭시켰다. SmaI으로 처리한 pTZ19R에 ligation시킨 후 염기서열을 밝히고자 sequencing하였다. 각각의 prototype virus와 비교하여 상동성을 밝혔다. 두 바이러스에 대한 cDNA probe를 제작하여 Southern blot hybridization을 실시하였다. TGEV의 경우 백신주인 P45와 병독주인 Miller strain을 사용하였다. cDNA를 증폭시키기 위해 N1/N1R과 N2/N2R 두 가지 primer를 이용한 결과, N1/N1R primer의 경우 586bp 크기의 PCR product를 얻을 수 있었고, N2/N2R primers로 582bp의 cDNA를 증폭시킬 수 있었다. PEDV 실험을 위하여 PED 임상 증상을 나타내는 분변을 이용하여 RT-PCR을 실시하였다. P2/P2R primer로 753bp의 PCR product를 얻을 수 있었다. TGEV의 두 가지 strain의 N protein gene을 sequencing하여 prototype인 Purdue strain과 염기서열 상동성을 조사한 결과, 97%이상의 높은 homology를 나타내었다. PED-V 역시 N protein gene을 sequencing하여 CV777과 염기서열 상동성을 조사한 결과 97%이상의 homology로 PEDV임을 알 수 있었다. TGEV와 PEDV의 염기서열을 비교한 결과 29%의 낮은 homology를 관찰할 수 있었다. 두 가지 바이러스의 N protein gene에 대한 cDNA probe를 제작하여 Southern blot hybridization을 한 결과, 각 바이러스에 매우 특이적 반응을 나타내었다.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.66 no.4
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• pp.318-325
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• 2021

The objective of this study was to identify the effect of zebularine, a DNA methylation inhibitor, on the chromosomal association between homoeologous chromosomes in the wheat genetic background. Zebularine at a final concentration of 10 µM was used to treat the spikes of the double monosomic wheat addition line (DMA) with one Leymus mollis chromosome and one Leymus racemosus chromosome, both of which were in a homoeologous relationship. In late prophase, zebularine led to chromosome breakage in the Leymus homoeologous chromosomes. Chromosome breakage caused an increase in the frequency of chromosomal associations between the Leymus homoeologous chromosomes. Ordinary DMA showed 65 cells (35.3%) with chromosomal associations and 119 cells (64.7%) with no association, whereas treated DMA showed 102 cells (60.0%) with chromosomal associations and 67 cells (39.4%) with no association. In diakinesis, the Leymus bivalent showed a chromosomal association in the whole euchromatic region. In metaphase, the Leymus bivalent showed association in the whole chromosomal region, unlike other Leymus bivalents with partial chromosomal association. Chromosomal association by chromosome breakage occurred not only between Leymus chromosomes but also between Leymus and wheat chromosomes. The frequency of other chromosomal association (such as fusion and insert) was increased. Chromosome breakage by zebularine treatment is a useful method at the chromosome level as the spores with others are hereditary stable, although the homologous index (h) was not significantly different between ordinary DMA and treated DMA. It is necessary to study how to control zebularine treatment with a more stable concentration for chromosome breakage during meiosis.

• Korean Journal of Plant Resources
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• v.37 no.2
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• pp.214-224
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• 2024

In this study, the growth, physical characteristics of fruit and soil chemistry were investigated by grouping S. thea populations. The soil for each group was sandy loam or sandy loam, and it was analyzed to be slightly acidic to neutral, with a pH ranging from 5.6 to 7.0. Additionally, the leaf size was highest in the G7 group, and overall fruit growth was highest in the G1 group, confirming differences between the groups. The sweetness ranged from 16.8° Brix to 12.3° Brix, indicating a higher sweetness compared to Vaccinium oldhamill, blueberry, and blackberry. The correlation analysis between soil chemistry and S. thea growth characteristics revealed a significant negative correlation between calcium (Ca) and fruit growth characteristics. Furthermore, a significant positive correlation was observed between sodium (Na) and fruit acidity, as well as between cation exchange capacity (CEC) and fruit hardness.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.189-194
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• 2008

The phospholipase B (PLB) families are enzymes sharing phospholipase (PL), lysophospholipase (LPL) and lysophospholipase-transacylase (LPTA) activities. In this study, we report the putative gene encoding phospholipase B (plbA) containing lipase motifs was cloned for the first time from the filamentous fungus, Aspergillus nidulans. plbA was isolated from A. nidulans genomic DNA library using a PCR-amplified probe, which is designed on the basis of sequence information derived from the conserved lipase regions of various PLBs. The deduced product of plbA is of 626 amino acids. From the assigned sequence, PlbA showed 72% identity with Penicillium notatum PLB but have low similarity with phospholipase A of other organisms.