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Construction of an Agroinfectious Clone of a Korean Isolate of Sweet Potato Symptomless Virus 1 and Comparison of Its Infectivity According to Agrobacterium tumefaciens Strains in Nicotiana benthamiana

  • Phuong T. Ho;Hee-Seong Byun;Thuy T. B. Vo;Aamir Lal;Sukchan Lee;Eui-Joon Kil
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.255-264
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    • 2023
  • Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus Mastrevirus (family Geminiviridae) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into Nicotiana benthamiana using three Agrobacterium tumefaciens strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The A. tumefaciens strain LBA4404 was the most effective at transferring the SPSMV-1 genome to N. benthamiana. We confirmed the viral replication in N. benthamiana samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

The Study of Trnascriptional Regulated Gene, $hrp^{2+}$, in Yeast

  • Choi, In-Soon
    • Journal of Life Science
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    • v.11 no.2
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    • pp.111-115
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    • 2001
  • This study was designed to clone the SNF2/SW12 helicase-related genes from the fission yeast Schizosaccha-romyces pombe and thereafter to elucidate the common functions of the proteins in this family. The $hrp^{2+}$gene was cloned by polymerase chain reaction amplification using degenerative primers from conserved SNF2 motifs within the ERCC6 gene, which encodes a protein involved in DNA excision repair. Like other SNF2/SW12 family proteins, the deduced amino acid sequence of Hrp2 contains DNA-dependent ATPase/7 helicase domains as well as the chromodomain and the DNA binding domain. This configuration is similar to that of mCHD1 (mouse chromo-ATPase/helicase-DNA-dinding protein 1), suggesting that Hrp2 is a S. pombe homolog of mCHD1, which is thought to function in altering the chromatin structure to control the gene expression. To characterize the function of Hrp2, 4 Uracil-Hrp2 fusion protein, it was purified near homogeneity by affinity chromatography on $Ni^{2+}$-NTA agarose, DEAE-Sepharose ion exchange arid Sephacryl S-200 gel filtration chromatographies. The purified fusion protein exhibited DNA-dependent ATPase activity, which was stimulated by both double-stranded and single-stranded DNA. To determine the steady-state level of $hrp^{2+}$ transcripts during growth, cells were cultured in medium and collected at every 2hr to prepare total RNAs. The northern blot analysis showed that the level of $hrp^{2+}$ transcripts reached its maximum before the cells entered the exponential growth phase and then decreased gradually, This result implies that Hrp2 may be required at early stages of cell growth.h.

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A Search for Exoplanets around Northern Circumpolar Stars. IX. A Multi-Period Analysis of the M Giant HD 135438

  • Byeong-Cheol Lee;Jae-Rim Koo;Yeon-Ho Choi;Tae-Yang Bang;Beomdu Lim;Myeong-Gu Park;Gwanghui Jeong
    • Journal of The Korean Astronomical Society
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    • v.56 no.2
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    • pp.277-286
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    • 2023
  • It is difficult to distinguish the pure signal produced by an orbiting planetary companion around giant stars from other possible sources, such as stellar spots, pulsations, or certain activities. Since 2003, we have obtained radial (RV) data from evolved stars using the high-resolution, fiber-fed Bohyunsan Observatory Echelle Spectrograph (BOES) at the Bohyunsan Optical Astronomy Observatory (BOAO). Here, we report the results of RV variations in the binary star HD 135438. We found two significant periods: 494.98 d with eccentricity of 0.23 and 8494.1 d with eccentricity of 0.83. Considering orbital stability, it is impossible to have two companions in such close orbits with high eccentricity. To determine the nature of the changes in the RV variability, we analyzed indicators of stellar spot and stellar chromospheric activity to find that there are no signals related to the significant period of 494.98 d. However, we calculated the upper limits of rotation period of the rotational velocity and found this to be 478-536 d. One possible interpretation is that this may be closely related to the rotational modulation of an orbital inclination at 67-90 degrees. The other signal corresponding to the period of 8494.1 d is probably associated with a stellar companion orbiting the giant star. A Markov Chain Monte Carlo (MCMC) simulation considering a single companion indicates that HD 135438 system hosts a stellar companion with 0.57+0.017 -0.017 M with an orbital period of 8498 d.

Implementation of a Deep Learning-Based Computer-Aided Detection System for the Interpretation of Chest Radiographs in Patients Suspected for COVID-19

  • Eui Jin Hwang;Hyungjin Kim;Soon Ho Yoon;Jin Mo Goo;Chang Min Park
    • Korean Journal of Radiology
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    • v.21 no.10
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    • pp.1150-1160
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    • 2020
  • Objective: To describe the experience of implementing a deep learning-based computer-aided detection (CAD) system for the interpretation of chest X-ray radiographs (CXR) of suspected coronavirus disease (COVID-19) patients and investigate the diagnostic performance of CXR interpretation with CAD assistance. Materials and Methods: In this single-center retrospective study, initial CXR of patients with suspected or confirmed COVID-19 were investigated. A commercialized deep learning-based CAD system that can identify various abnormalities on CXR was implemented for the interpretation of CXR in daily practice. The diagnostic performance of radiologists with CAD assistance were evaluated based on two different reference standards: 1) real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) results for COVID-19 and 2) pulmonary abnormality suggesting pneumonia on chest CT. The turnaround times (TATs) of radiology reports for CXR and rRT-PCR results were also evaluated. Results: Among 332 patients (male:female, 173:159; mean age, 57 years) with available rRT-PCR results, 16 patients (4.8%) were diagnosed with COVID-19. Using CXR, radiologists with CAD assistance identified rRT-PCR positive COVID-19 patients with sensitivity and specificity of 68.8% and 66.7%, respectively. Among 119 patients (male:female, 75:44; mean age, 69 years) with available chest CTs, radiologists assisted by CAD reported pneumonia on CXR with a sensitivity of 81.5% and a specificity of 72.3%. The TATs of CXR reports were significantly shorter than those of rRT-PCR results (median 51 vs. 507 minutes; p < 0.001). Conclusion: Radiologists with CAD assistance could identify patients with rRT-PCR-positive COVID-19 or pneumonia on CXR with a reasonably acceptable performance. In patients suspected with COVID-19, CXR had much faster TATs than rRT-PCRs.

Occurrence and Multiplex PCR Detection of Citrus Yellow Vein Clearing Virus in Korea

  • Taemin Jin;Ji-Kwang Kim;Hee-Seong Byun;Hong-Soo Choi;Byeongjin Cha;Hae-Ryun Kwak;Mikyeong Kim
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.125-138
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    • 2024
  • Citrus yellow vein clearing virus (CYVCV) is a member of the Alphaflexiviridae family that causes yellow vein clearing symptoms on citrus leaves. A total of 118 leaf samples from nine regions of six provinces in Korea were collected from various citrus species in 2020 and 2021. Viral diagnosis using next-generation sequencing and reverse transcription polymerase chain reaction (RT-PCR) identified four viruses: citrus tristeza virus, citrus leaf blotch virus, citrus vein enation virus, and CYVCV. A CYVCV incidence of 9.3% was observed in six host plants, including calamansi, kumquat, Persian lime, and Eureka lemon. Among the citrus infected by CYVCV, only three samples showed a single infection; the other showed a mixed infection with other viruses. Eureka lemon and Persian lime exhibited yellow vein clearing, leaf distortion, and water-soak symptom underside of the leaves, while the other hosts showed only yellowing symptoms on the leaves. The complete genome sequences were obtained from five CYVCV isolates. Comparison of the isolates reported from the different geographical regions and hosts revealed the high sequence identity (95.2% to 98.8%). Phylogenetic analysis indicated that all the five isolates from Korea were clustered into same clade but were not distinctly apart from isolates from China, Pakistan, India, and Türkiye. To develop an efficient diagnosis system for the four viruses, a simultaneous detection method was constructed using multiplex RT-PCR. Sensitivity evaluation, simplex RT-PCR, and stability testing were conducted to verify the multiplex RT-PCR system developed in this study. This information will be useful for developing effective disease management strategies for citrus growers in Korea.

Translocator Protein (18 kDa) Polymorphism (rs6971) in the Korean Population

  • Hyon Lee;Young Noh;Woo Ram Kim;Ha-Eun Seo;Hyeon-Mi Park
    • Dementia and Neurocognitive Disorders
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    • v.21 no.2
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    • pp.71-78
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    • 2022
  • Background and Purpose: The expression of the 18-kDA mitochondrial translocator protein (TSPO) in the brain is an attractive target to study neuroinflammation. However, the binding properties of TSPO ligands are reportedly dependent on genetic polymorphism of the TSPO gene (rs6971). The objective of this study is to investigate the rs6971 gene polymorphism in the Korean population. Methods: We performed genetic testing on 109 subjects including patients with mild cognitive impairment, Alzheimer's disease (AD) dementia, non-AD dementia, and cognitively unimpaired participants. Magnetic resonance imaging scans and detailed neuropsychological tests were also performed, and 29 participants underwent 18F-DPA714 PET scans. Exon 4 of the TSPO gene containing the polymorphism rs6971 (Ala or Thr at position 147) was polymerase chain reaction amplified and sequenced using the Sanger method. The identified rs6971 genotype codes (C/C, C/T, or T/T) of the TSPO protein generated high-, mixed-, or low-affinity binding phenotypes (HABs, MABs, and LABs), respectively. Results: We found that 96.3% of the study subjects were HAB (105 out of 109 subjects), and 3.7% of the subjects were MAB (4 out of 109 subjects). 18F-DPA-714 PET scans showed nonspecific binding to the thalamus and brainstem, and increased tracer uptake throughout the cortex in cognitively impaired patients. The participant with the MAB polymorphism had a higher DPA714 signal throughout the cortex. Conclusions: The majority of Koreans are HAB (aprox. 96%). Therefore, the polymorphism of the rs6971 gene would have a smaller impact on the availability of second-generation TSPO PET tracers.

Association analysis of polymorphisms in six keratin genes with wool traits in sheep

  • Sulayman, Ablat;Tursun, Mahira;Sulaiman, Yiming;Huang, Xixia;Tian, Kechuan;Tian, Yuezhen;Xu, Xinming;Fu, Xuefeng;Mamat, Amat;Tulafu, Hanikezi
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.775-783
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    • 2018
  • Objective: The purpose of this study was to investigate the genetic effects of six keratin (KRT) genes on the wool traits of 418 Chinese Merino (Xinjiang type) (CMXT) individuals. Methods: To explore the effects and association of six KRT genes on sheep wool traits, The polymerase chain reaction-based single-strand conformation polymorphism (PCR-SSCP), DNA sequencing, and the gene pyramiding effect methods were used. Results: We report 20 mutation sites (single-nucleotide polymorphisms) within the six KRT genes, in which twelve induced silent mutations; five induced missense mutations and resulted in $Ile{\rightarrow}Thr$, $Glu{\rightarrow}Asp$, $Gly{\rightarrow}Ala$, $Ala{\rightarrow}Ser$, $Se{\rightarrow}His$; two were nonsense mutations and one was a same-sense mutation. Association analysis showed that two genotypes of the KRT31 gene were significantly associated with fiber diameter (p<0.05); three genotypes of the KRT36 gene were significantly associated with wool fineness score and fiber diameter (p<0.05), three genotypes of the KRT38 gene were significantly associated with the number of crimps (p<0.05); and three genotypes of the KRT85 gene were significantly associated with wool crimps score, body size, and fiber diameter (p<0.05). Analysis of the gene pyramiding effect between the different genotypes of the gene loci KRT36, KRT38, and KRT85, each genotype in a gene locus was combined with all the genotypes of another two gene loci and formed the different three loci combinations, indicated a total of 26 types of possible combined genotypes in the analyzed population. Compared with the other combined genotypes, the combinations CC-GG-II, CC-HH-IJ, CC-HH-JJ, DD-HH-JJ, CC-GH-IJ, and CC-GH-JJ at gene loci KRT36, KRT38, and KRT85, respectively, had a greater effect on wool traits (p<0.05). Conclusion: Our results indicate that the mutation loci of KRT31, KRT36, KRT38, and KRT85 genes, as well as the combinations at gene loci KRT36, KRT38, and KRT85 in CMXT have significant effects on wool traits, suggesting that these genes are important candidate genes for wool traits, which will contribute to sheep breeding and provide a molecular basis for improved wool quality in sheep.

Intensified Low-Temperature Fischer-Tropsch Synthesis Using Microchannel Reactor Block : A Computational Fluid Dynamics Simulation Study (마이크로채널 반응기를 이용한 강화된 저온 피셔-트롭쉬 합성반응의 전산유체역학적 해석)

  • Kshetrimatum, Krishnadash S.;Na, Jonggeol;Park, Seongho;Jung, Ikhwan;Lee, Yongkyu;Han, Chonghun
    • Journal of the Korean Institute of Gas
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    • v.21 no.4
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    • pp.92-102
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    • 2017
  • Fischer-Tropsch synthesis reaction converts syngas (mixture of CO and H2) to valuable hydrocarbon products. Simulation of low temperature Fischer -Tropsch Synthesis reaction and heat transfer at intensified process condition using catalyst filled single and multichannel microchannel reactor is considered. Single channel model simulation indicated potential for process intensification (higher GHSV of $30000hr^{-1}$ in presence of theoretical Cobalt based super-active catalyst) while still achieving CO conversion greater than ~65% and $C_{5+}$ selectivity greater than ~74%. Conjugate heat transfer simulation with multichannel reactor block models considering three different combinations of reactor configuration and coolant type predicted ${\Delta}T_{max}$ equal to 23 K for cross-flow configuration with wall boiling coolant, 15 K for co-current flow configuration with subcooled coolant, and 13 K for co-current flow configuration with wall boiling coolant. In the range of temperature maintained (498 - 521 K), chain growth probability calculated is desirable for low-temperature Fisher-Tropsch Synthesis.

Species Diversity of the Dinoflagellate Genus Alexandrium in the Coastal Waters of Korea during Summer 2013 (2013년 하계 국내 연안에서 출현하는 Alexandrium 속 와편모류의 종 다양성)

  • KIM, JAE SEONG;PARK, KYUNG WOO;YOUN, SEOK HYUN;LIM, WEOL AE;YOO, YEONG DU;SEONG, KEYONG AH;YIH, WON HO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.21 no.4
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    • pp.158-170
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    • 2016
  • We investigated the occurrence of the dinoflagellate genus Alexandrium in the nineteen Korean coastal sites from July to October 2013. Alexandrium-like planktonic cells were microscopically observed only in four out of the 19 sampling sites. From the samples containing Alexandrium-like cells 22 clonal cultures of Alexandrium species were established by single cell or single chain isolation method. Taxonomic identity of the 4 different strains ascertained by the robust analyses of morphological and molecular genetic characteristics were confirmed to be A. catenella, A. affine, A. fraterculus and an unidentified Alexandrium sp. for which strain WEB-Alex-01 was assigned. It was ascertained that in spite of hot summer diverse Alexandrium species attaining up to four were distributed in the study area, in contrast with the long empirical recognition that the emergence of Alexandrium species is restricted to cooler seasons like spring or autumn in Korean coastal waters. Morphology and genetic characteristics of Alexandrium sp. strain WEB-Alex-01 are different from any other previously reported Alexandrium species from Korean seas, which implies that further studies on taxonomic, physiological, ecological and toxicological properties of the newly recorded Alexandrium species are needed.

Development of the Duplex PCR Method of Identifying Trachurus japonicus and Trachurus novaezelandiae (다중 PCR 분석법을 이용한 전갱이속 어종의 신속한 종판별 분석법 개발)

  • Park, Yeon Jung;Lee, Mi Nan;Kim, Eun Mi;Noh, Eun Soo;Noh, Jae Koo;Park, Jung Youn;Kang, Jung-Ha
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1062-1067
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    • 2018
  • Reliable labeling of fish products can reassure consumers regarding the identity and quality of seafoods. Therefore, techniques that can identify adulteration or mislabeling are valuable. To rapidly identify two Trachurus species, Trachurus japonicus and Trachurus novaezelandiae, a highly efficient, rapid, duplex polymerase chain reaction (PCR) having two species-specific primers simultaneously was identified. This species-specific primer focused on a single nucleotide mismatch in the 3'-terminal base of a primer designed in the mitochondrial cytochrome c oxidase (COI) subunit I DNA. To optimize the duplex PCR condition, gradient PCR reactions were conducted to determine the primer annealing temperature and the primer concentration. The PCR's product was observed on the gel, suggesting that DNA molecules may be useful in differentiating the two species. The length of the amplification fragments were 103 bp for Trachurus japonicus and 214 bp for Trachurus novaezelandiae, which, along with the species-specific primer visualized by agarose gel electrophoresis, enabled accurate distinction of the species of the Trachurus genus. These results indicate that the duplex PCR, which has a species-specific primer based on single nucleotide polymorphism (SNP), can be useful for rapidly differentiating the two species of Trachurus. This duplex PCR analysis is simple, rapid, and reliable, and could be beneficial to protecting consumers' rights.