• Parasites, Hosts and Diseases
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• 제51권2호
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• pp.237-241
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• 2013

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from $10^{-1}$ to $10^{-5}ng/{\mu}l$ for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of $63^{\circ}C$ by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha ($EF1{\alpha}$) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

• The Plant Pathology Journal
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• 제22권2호
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• pp.168-173
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• 2006

To develop the diagnostic method for the viral infection in apple, the partial genes corresponding to the N-terminal region of RNA polymerase of Apple stem grooving virus (ASGV) and coat protein of Apple chlorotic leaf spot virus (ACLSV) were characterized from the infected apple cultivars in Korea. Based on the nucleotide sequences of the characterized partial genes, the virus gene-specific primers were designed for the detection of ASGV and ACLSV infected in species of Malus. The RT-PCR using the primers for the genes of ASGV and ACLSV successfully gave rise to 404 and 566 bp DNA fragments, respectively. Using those viral gene-specific primers, the multiplex RT-PCR assays were also established to diagnose the mixed infection by ASGV and ACLSV simultaneously. Furthermore, the control primers, which have to be included for the RT-PCR as an internal control, were designed using the nucleotide sequence of the gene encoding elongation factor $1{\alpha}(EF1{\alpha})$. This multiplex RT-PCR including the control primers provides more reliable, rapid and sensitive assay for the detection of ASGV and ACLSV infected in Korean apple cultivars.

• The Plant Pathology Journal
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• 제33권3호
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• pp.249-263
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• 2017

This study aims to examine the potential reasons for the current prevalence of the fusarium wilt in the oriental melon. Twenty-seven Fusarium isolates obtained from oriental melon greenhouses in 2010-2011 were identified morphologically and by analysis of elongation factor-1 alpha gene (EF-$1{\alpha}$) and internal transcribed spacer (ITS) rDNA sequences as 6 Fusarium species (8 isolates of F. oxysporum, 8 F. commune, 5 F. proliferatum, 3 F. equiseti, 2 F. delphinoides, and 1 F. andiyazi), which were classified as same into 6 EF-$1{\alpha}$ sequence-based phylogenetic clades. Pathogenicity of the Fusarium isolates on the oriental melon was highest in F. proliferatum, next in F. oxysporum and F. andiyazi, and lowest in the other Fusarium species tested, suggesting F. proliferatum and F. oxysporum were major pathogens of the oriental melon, inducing stem rots and vascular wilts, respectively. Oriental melon and watermelon were more susceptible to F. oxysporum than shintosa and cucumber; and cucumber was most, oriental melon and watermelon, medially, and shintosa was least susceptible to F. proliferatum, whose virulence varied among and within their phylogenetic subclades. Severe root-knot galls were formed on all the crops infected with Meloidogyne incognita; however, little indication of vascular wilts or stem and/or root rots was shown by the nematode infection. These results suggest the current fungal disease in the oriental melon may be rarely due to virulence changes of the fusarium wilt pathogen and the direct cause of the severe root-knot nematode infection, but may be potentially from other Fusarium pathogen infection that produces seemingly wilting caused by severe stem rotting.

• Food Science and Biotechnology
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• 제14권6호
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• pp.854-859
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• 2005

Although action modes of equol and genistein have been extensively studied, their precise roles in tumor cells remain elusive. To address possible effects of these compounds on protein expression in mammary tumor cells, proteins modulated in MCF-7 mammary tumor cells when incubated in absence and presence of 10 uM equol or genistein were identified through 2-dimensional gel electrophoresis, MALDI-TOF MS/MS, and NCBInr database search using Mascot software. Most proteins differentially expressed in MCF-7 cells after treatment with 10 uM genistein or equol were identified as being the same. Exposure to both compounds caused decreased cellular expression of RNA-binding protein regulatory subunit and oncogene DJ1 tubulin beta-1 chain, and increased expression of heterogeneous ribonucleoproteins F and L, KH-type splicing regulatory protein, and translation elongation factor EF-Tu precursor. Genistein and equol at dose used in this study showed common action mechanism.

• Mycobiology
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• 제48권6호
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• pp.495-500
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• 2020

Leaf spot on lemon balm is frequently observed in Korea, causing considerable damage to crops. In 2014 and 2015, the occurrence of leaf spot was observed in several production greenhouses at Suwon, Gongju, and Namwon in Korea. Symptoms on lower leaves initially developed as small, distinct, discolored lesions, which enlarged progressively turning into dark brown, angular spots surrounded by purplish-brown margins. Based on the morphological characteristics and sequence analysis of actin (ACT), translation elongation factor 1-alpha (EF-1α), internal transcribed spacer (ITS), 28S nrDNA (LSU), and RNA polymerase II second largest subunit (RPB2), the fungus associated with the lemon balm leaf spot was determined as Septoria melissae. To the best of our knowledge, this is the first report of lemon balm leaf spot caused by S. melissae in Asia as well as in Korea.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.80.1-80
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• 2003

Several plant and microbial genes that could confer disease resistance in transgenic rice plants are being cloned and characterized. We are currently constructing transgenic rice lines that overexpress the gene products, such as a galactinol synthase, a defensin, and a bacterial ACC deaminase. Subtractive hybridization of a rice cDNA library constructed from the Xanthomonas oryzae-infected ice leaves resulted in isolation of many inducible cDNA clones including a elongation factor EF2, a oryzain alpha, a catalase, a aldehyde dehydrogenase, a S-adenosylmethionine synthetase, a caffeic acid O-methyltransferase, a glyceraldehyde-3-phosphate dehydrogenase, a light-regulated protein, nKY transcription factors, and a nucleotide diphosphate kinase. Some genes among those may be useful genetic sources for construction of disease resistant transgenic rice. Full lengths of the rice OsFIERG and a rice oryzain genomic clones were cloned, and serial deletion fragments of the promoter regions of these genes were fused with GUS reporter gene in pCAMBIA1201, respectively. Promoter activities of these constructs will be examined upon various stresses and Pathogen infections to obtain the pathogen specific inducible-promoter. This work was supported by a grant from BioGreen 21 Program, Rural Development Administration, Republic of Korea.

• Bulletin of the Korean Chemical Society
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• 제22권12호
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• pp.1361-1365
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• 2001

Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.1003-1010
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• 2008

The objective of the present study was to identify differences in the expression levels of liver proteins between healthy and ketotic cows, establish a liver metabolic interrelationship of ketosis and elucidate the metabolic characteristics of the liver during ketosis. Liver samples from 8 healthy multiparous Hostein cows and 8 ketotic cows were pooled by health status and the proteins were separated by two-dimensional-electrophoresis (2D-E). Statistical analysis of gels was performed using PDQuest software 8.0. The differences in the expression levels of liver proteins (p<0.05) between ketotic and healthy cows were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry. Five enzymes/proteins were identified as being differentially expressed in the livers of ketotic cows: expression of 3-hydroxyacyl-CoA dehydrogenase type-2 (HCDH), acetyl-coenzyme A acetyltransferase 2 (ACAT) and elongation factor Tu (EF-Tu) were down-regulated, whereas that of alpha-enolase and creatine kinase were up-regulated. On the basis of this evidence, it could be presumed that the decreased expression of HCDH, which is caused by high concentrations of acetyl-CoA in hepatic cells, in the livers of ketotic cows, implies reduced fatty acid ??oxidation. The resultant high concentrations of acetyl-CoA and acetoacetyl CoA would depress the level of ACAT and generate more ??hydroxybutyric acid; high concentrations of acetyl-CoA would also accelerate the Krebs Cycle and produce more ATP, which is stored as phosphocreatine, as a consequence of increased expression of creatine kinase. The low expression level of elongation factor Tu in the livers of ketotic cows indicates decreased levels of protein synthesis due to the limited availability of amino acids, because the most glucogenic amino acids sustain the glyconeogenesis pathway; thus increasing the level of alpha-enolase. Decreased protein synthesis also promotes the conversion of amino acids to oxaloacetate, which drives the Krebs Cycle under conditions of high levels of acetyl-CoA. It is concluded that the livers of ketotic cows possess high concentrations of acetyl-CoA, which through negative feedback inhibited fatty acid oxidation; show decreased fatty acid oxidation, ketogenesis and protein synthesis; and increased gluconeogenesis and energy production.

• 식물병연구
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• 제26권2호
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• pp.79-87
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• 2020

Fusarium oxysporum f. sp. fragariae (Fof) 에 의한 딸기 시들음병은 국내 딸기재배에서 가장 중요한 병해 중 하나이다. 국내 발생하는 Fof의 특성을 분석하고자 시들음병균의 유전적 다양성, 병원성과 살균제 반응을 조사하였다. 분리균은 Fo080701를 제외한 모든 균주에서 Fof 특이적 primer에 증폭되었다. 분리균의 nuclear ribosomal intergenic spacer region과 EF-1α sequences 분석 결과 3개의 lineage를 형성하였다. 대부분의 분리균은 lineage 1에 속하였으며 lineage 3에 3개 균주와 lineage 2에 1개 균주가 포함되었다. 분리된 모든 균주는 설향품종에 병원성을 보였다. Prochloraz는 DNA lineage 2에 속하는 Fo080701균주를 제외하곤 시들음병균의 EC₅₀값이 0.02-0.1 ㎍/ml로 낮은 농도에서 효과적으로 균사 생장을 억제하였다. Metconazole의 EC₅₀값도 0.04-0.22 ㎍/ml로 prochloraz와 비슷한 억제 효과를 보였다. Pyraclostrobin의 EC₅₀값은 0.23-168.01 ㎍/ml로 균주에 따라 차이가 컸다. 딸기 재배포장에서 boscalid+fludioxonil, fluxapyroxad+pyraclostrobin, prochloraz manganese이 딸기 시들음병 방제에 효과적이었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제26권2호
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• pp.95-112
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• 2013

As a first step toward understanding the divergence and relationships of the Nymphalidae (Lepidoptera: Papilionoidea) occurring in South Korea, cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-$1{\alpha}$ (EF-$1{\alpha}$) that comprise 3,501-3,716 bp were either sequenced (55 species) or the sequences were obtained from GenBank (23 species). The concatenated sequence divergence of six nymphalid subfamilies ranked in the following order: Danainae (10.3%), Satyrinae (9.5%), Limenitidinae (8.0%), Apaturinae (7.0%), Nymphalinae (6.7%), and Heliconiinae (6.2%). As has been reported in previous large scale international studies, the subfamilial relationships of (((((Limenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Satyrinae) + Libytheinae) + Danainae) were also confirmed, except for the switched positions between Danainae and Libytheinae, and supported all subfamilies and tribe monophylies. Unlikely consistent phylogenetic relationships among genera within the majority of tribes in Nymphalidae, a conflicting relationship within the subfamily Apaturinae was obvious, presenting Apatura as sister to either Mimathyma or (Mimathyma + (Sephisa + (Hestina + Sasakia))), and both of these relationships are unconventional. Within the subfamily Limenitidinae, the genus Neptis was consistently revealed as a paraphyletic with respect to the genus Aldania, requiring further taxonomic investigation of the genus. Although limited, current sequence information and phylogenetic relationships are expected to be helpful for further studies.