• Korean Journal of Agricultural Science
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• v.37 no.3
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• pp.367-371
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• 2010

Farnesyl diphosphate synthase (FPS) catalyzes the biosynthesis of farnesyl diphosphate, a precursor for many important terpenoid products. A cDNA encoding FPS was first isolated from Allium sativum (AsFPS) using rapid amplification of cDNA ends (RACE) PCR. The sequence of AsFPS contains an open reading frame encoding a protein of 341 amino acids with a predicted molecular mass of 39.61 kDa. Alignment of AsFPS deduced amino acid revealed high identities with other plants ranging from 79% to 85% and showed 2-high conserved aspartate-rich motifs known to be important for FPS activity. Furthermore, AsFPS expression was stronger in the green organs such as bulbils, scapes, leaves, stems, but weaker in bulbs and roots than on-green organs of A. sativum.

• Genomics & Informatics
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• v.3 no.1
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• pp.24-29
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• 2005

TRPV2 is a non-specific cation channel expressed in sensory neurons, and activated by noxious heat. Particularly, TRPV2 has six transmembrane domains and three ankyrin repeats. TRPV2 has been cloned from various species such as human, rat, and mouse. Oocytes of Xenopus laevis - an African clawed frog ­have been widely used for decades in characterization of various receptors and ion channels. The functional property of rat TRPV2 was also identified by this oocyte expression system. However, no TRPV2 orthologue of Xenopus laevis has been reported so far. Hence, we have focused to clone a TRPV2 orthologue of Xenopus laevis with the aid of bioinformatic tools. Because the genome sequence of Xenopus laevis is not available until now, a genome sequence of Xenopus tropicalis - a close relative species of Xenopus laevis - was used. After a number of bioinformatic searches in silico, a predicted full-length sequence of TRPV2 orthologue of Xenopus tropicalis was found. Based on this predicted sequence, various approaches such as RT-PCR and 5' -RACE technique were applied to clone a full length of Xenopus laevis TRV2. Consequently, a full-length Xenopus laevis TRPV2 was cloned from heart cDNA.

• BMB Reports
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• v.39 no.5
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• pp.607-617
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• 2006

A novel calcium-dependent protein kinase gene (designated as IiCPK2) was cloned from tetraploid Isatis indigotica. The full-length cDNA of IiCPK2 was 2585 bp long with an open reading frame (ORF) of 1878 bp encoding a polypeptide of 625 amino acid residues. The predicted IiCPK2 polypeptide included three domains: a kinase domain, a junction domain (or autoinhibitory region), and a C-terminal calmodulin-like domain (or calcium-binding domain), which presented a typical structure of plant CDPKs. Further analysis of IiCPK2 genomic DNA revealed that it contained 7 exons, 6 introns and the length of most exons was highly conserved. Semi-quantitative RT-PCR revealed that the expression of IiCPK2 in root, stem and leaf were much higher in tetraploid sample than that in diploid progenitor. Further expression analysis revealed that gibberellin ($GA_3$), NaCl and cold treatments could up-regulate the IiCPK2 transcription. All our findings suggest that IiCPK2 might participate in the cold, high salinity and GA3 responsive pathways.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.4
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• pp.319-322
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• 2002

Soybean cyst nematode (Heterodera glycines Ichinohe; SCN) is an important soybean pest and the use of resistant cultivars is the effective method to reduce or eliminate SCN damage. However, breeding for SCN resistance is difficult and expensive by the oligogenic nature of the resistance and genetic variability in the pathogen. Fortunately, SCN resistance loci, rhg1 and Rhg4 are generally accepted as a necessity for the development of resistant genotypes using any source of resistance. In this study, resistance of 44 Korean soybean cultivars to SCN was tested using two molecular markers. Seonheukkong and Pokwangkong were the homozygous to rhg1 locus. Seven cultivars were susceptible to SCN based on Satt309 marker linked rhg1 locus. All Korean cultivars estimated in this study were recessive homozygous to Rhg4 locus and were susceptible in the PCR reaction using primer 548/563 linked to the Rhg4 locus conferring resistance to SCN race 3. Among 44 cultivars estimated, seven cultivars were susceptible to SCN in both Satt309 and primer 548/563 markers. Based on both Satt309 and primer 548/563 markers, there is no resistant cultivar to SCN in Korea. Therefore, SCN resistant cultivars need to be developed in the future. These two markers can be used for improving SCN resistant cultivars.

• Korean Journal of Breeding Science
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• v.42 no.2
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• pp.139-143
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• 2010

Late blight caused by Phytophthora infestans is historically a serious epidemic disease in potato and tomato cultivations. Accession L3708 (Solanum pimpinellifolium), a new source for late blight resistance was identified in AVRDC, and carries the resistance gene, Ph-3, incompatible to P. infestans race 3. The AFLP markers linked to Ph-3 were previously developed from the L3708 accession (Chunwongse et al. 2002). To facilitate tomato breeding with the Ph-3 gene, an attempt was made to convert AFLP markers to sequence-characterized amplified region (SCAR) markers. Among 6 AFLP markers, only one AFLP marker, L87, was successfully converted to SCAR marker. The resistance-specific 230 bp AFLP fragment was cloned and sequenced, and the PCR primer amplifying a 123 bp fragment was designed. This SCAR marker could discriminate resistant and susceptible individuals with high stringency. The developed SCAR marker could be used for the marker assisted-selection in tomato breeding programs.

• Journal of The Korean Society of Grassland and Forage Science
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• v.40 no.4
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• pp.285-290
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• 2020

To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.113.1-113
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• 2003

A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.), and its entire genome RNA sequence was determined. in Korea using cDNA library and RACE methods. The genome of HVX encodes five open reading frames coding for viral replicase, triple gene block (TGB), and viral coat protein (CP) from the 5'to 3' ends, which is a typical genome structure of potexviruses. The 3-terminal region of the virus includes the TGBI (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3-nontranslated region (NTR). The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100％ and 98.9％ identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50％ nucleotide identical to other sequenced potexviruses. This is the first report of complete genome sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.4
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• pp.314-326
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• 2020

This study was carried out to develop a resistant variety against the K3a race of bacterial blight, Xanthomonas oryzae pv. oryzae, through expansion and pyramiding of resistance genes. To develop an elite bacterial blight-resistant cultivar, the breeding process and bacterial blight resistance reactions in advanced backcross lines (ABLs) were analyzed. ABLs21 which contain Xa3 and Xa21, were developed by double backcrossing japonica cultivar Hwanggeumnuri, which has bacterial blight resistant Xa3 gene, and indica variety IRBB21, which havs Xa21 gene, followed by disease resistance bioassay and marker-assisted selection. The resistance genes of ABLs21 were amplified by PCR with the molecular markers 9643.T4 (Xa3) and U1/I1 (Xa21). Hwanggeumnuri and IRBB3 showed resistance reactions against K1, K2, and K3 races, and a susceptible reaction against K3a, K4, and K5 races. IRBB21 showed resistance reactions against K2, K3, K3a, K4 and K5 races, and a susceptible reaction against K1 race. Hwanggeumnuri showed susceptible reactions at the seedling, tillering and adult stages (all stages), whereas ABL21-1 showed moderate resistance at the tillering stage. ABL21-1 showed stable resistance against 18 isolates of K3a race, and the lesion length was shorter than that of the donor parents. In cluster analysis, the HB4032 isolate showed the highest pathogenicity among the 18 isolates. The molecular marker polymorphisms and average substituted chromosome segment lengths of ABLs21 were 63.2 % and 86.1 cM, respectively. Insertion of the donor chromosomal segments occurred in the predicted region of the Xa21 gene of ABLs21.

• Journal of Life Science
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• v.21 no.8
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• pp.1156-1162
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• 2011

Equine coital exanthema caused by equine herpesvirus type 3 (EHV-3) is a venereal disease which seriously drops horse reproduction rates. Here, we isolated EHV-3 from infected horses and investigated their biological characteristics. Initial cytopathic effects such as rounding of cells were detected 48 hours post infection of the virus into RK-13 cells. The infected cells were going to detach from the surface of culture flasks 72 hours post infection. The type of isolated viruses from swabbed samples was EHV-3 by PCR analysis. Glycoprotein G (gG) of isolated EHV-3 has a 99.25 percent similarity rate to that of EHV-3 334/74 control strain. The isolated EHV-3 was named Georo strain. Georo strain consisted of four major proteins including 145 kD, 60 kD, 45 kD and 40 kD, as shown by SDS-PAGE analysis. We hope the newly isolated Georo strain of EHV-3 can be used for studying various aspects of Korean equine coital exanthema.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.10
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• pp.1471-1477
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• 2017

Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.