• Title/Summary/Keyword: genetic analyses

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Molecular characterization and expression of a disintegrin and metalloproteinase with thrombospondin motifs 8 in chicken

  • Lee, Ra Ham;Lee, Seokhyun;Kim, Yu Ra;Kim, Sung-Jo;Lee, Hak-Kyo;Song, Ki-Duk
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1366-1372
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    • 2018
  • Objective: A disintegrin and metallopeptidase with thrombospondin motifs type 8 (ADAMTS8) is crucial for diverse physiological processes, such as inflammation, tissue morphogenesis, and tumorigenesis. The chicken ADAMTS8 (chADAMTS8) gene was differentially expressed in the kidney following exposure to different calcium concentrations, suggesting a pathological role of this protein in metabolic diseases. We aimed to examine the molecular characteristics of chADAMTS8 and analyze the gene-expression differences in response to toll-like receptor 3 (TLR3) stimulation. Methods: The ADAMTS8 mRNA and amino acid sequences of various species (chicken, duck, cow, mouse, rat, human, chimpanzee, pig, and horse) were retrieved from the Ensembl database and subjected to bioinformatics analyses. Reverse-transcription polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) experiments were performed with various chicken tissues and the chicken fibroblast DF-1 cell line, which was stimulated with polyinosinic-polycytidylic acid (poly[I:C]; a TLR3 ligand). Results: The chADAMTS8 gene was predicted to contain three thrombospondin type 1 (TSP1) domains, whose amino acid sequences shared homology among the different species, whereas sequences outside the TSP1 domains (especially the amino-terminal region) were very dif­ferent. Phylogenetic analysis revealed that chADAMTS8 is evolutionarily clustered in the same clade with that of the duck. chADAMTS8 mRNA was broadly expressed in chicken tissues, and the expression was significantly up-regulated in the DF-1 cells in response to poly(I:C) stimulation (p<0.05). These results showed that chADAMTS8 may be a target gene for TLR3 signaling. Conclusion: In this report, the genetic information of chADAMTS8 gene, its expression in chicken tissues, and chicken DF-1 cells under the stimulation of TLR3 were shown. The result suggests that chADAMTS8 expression may be induced by viral infection and correlated with TLR3-mediated signaling pathway. Further study of the function of chADAMTS8 during TLR3-dependent inflammation (which represents RNA viral infection) is needed and it will also be important to examine the molecular mechanisms during different regulation, depending on innate immune receptor activation.

Comparative Analysis of Acanthopanax senticosus Harms from Korea, China and Russia Based on the ITS Sequences of Nuclear Ribosomal DNA (ITS 염기서열분석에 의한 한국산, 중국산 및 러시아산 가시오갈피의 유연관계 분석)

  • Han Hyo-Shim;Kim Doo-Young;Lee Kab-Yeon;Park Wan-Geun;Cho In-Kyung;Jung Jae-Sung
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.54-58
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    • 2006
  • The genetic analyses of Acanthopanax senticosus Harms from Korea, China and Russia, were made by comparing the internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA. The ITS region of A. senticosus was amplified by polymerase chain reaction (PCR) using the universal primers and then directly sequenced. The length of the ITS region including 162 bp 5.85 rRNA gene ranged from 608 bp (for Korean and Chinese) to 611 bp (for Russian). The G+C content of ITS region were 60.20% for Korean and Chinese plants and 60.06% for Russian plants. Sequence comparisons indicated that ITS regions of A. senticosus from Korea and China were identical, whereas the ITS sequence of A. senticosus from Russia showed 99.2% homology with the plants from Korea. Variation in sequences were attributable to 5 bp substitution such as transversion or insertion events. These results suggested that A. senticosus Harms from Korea and China were closely related in phylogenetic relationship compared to Russian. In addition, A. senticosus Harms were more similar to Kalopanax pictus than A. sessiliflorus in their ITS sequences.

Characterization of the genomes of Aujeszky's disease virus isolated in Korea (국내분리 오제스키병 바이러스의 게놈 유전자 특성 분석)

  • Hyun, Bang-Hun;Kim, In-Joong;Pyo, Hyun-Mi;Cha, Sang-Ho;Park, Ji-Yeun;Song, Jae-Young;Cho, In-Soo;Yang, Chang-Bum;An, Soo-Hwan;Lee, Joong-Bok
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.45-57
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    • 2009
  • The molecular genetic characterization of Aujeszky's disease virus (ADV) Yangsan strain (ADVYS), a Korean isolate, was investigated by analyzing the electrophoresis patterns and the physical maps of the viral DNA digested with various endonucleases. To establish DNA library for ADV-YS, twelve major BamHI restricted segments were cloned. Each location of the segments in the ADV genome was determined by sequence comparison with the sequences reported in Genbank and those sequences of the both termini of the segments. Physical maps were constructed based on the electrophoresis patterns of the digested viral DNA by restriction endonuclease and the results of Southern blot analyses with various DIG labeled probes originated from those of enzyme restricted segments of virulent (Shope) and avirulent (Bartha) strain. Comparing ADV-YS with a standard strain of Kaplan in the maps of restriction enzymes, following major respects were identified: (i) disappearance of BamHI restriction site between the first and second BamHI segments, (ii) creation of the BamHI restriction site in the fifth segment, and (iii) generation of the BglII site in the unique short (US) region. The genome of ADV-YS also contains a type 2 herpesvirus DNA molecule (in which the US region only inverts itself relative to the unique longregion) like all other ADV strains except Norden strain(type3), analyzed up to date. The size of the ADV genome estimated from the sizes of the restriction enzyme fragments, was approximately 145.3 kb (BamHI) or 145.4 kb (BglII). BamHI enzyme cleavage patterns were compared among the five Korean ADV isolates: Yangsan, Yongin, Dangjin, Jincheon and Iksan strains. Difference either in the number or in the size of the DNA fragments, suspected regions of termini of IR and TR, could be detected among all five strains.

A LuxR-type Transcriptional Regulator, PsyR, Coordinates Regulation of Pathogenesis-related Genes in Pseudomonas syringae pv. tabaci (Pseudomonas syringae pv. tabaci 에서 LuxR-type 전사조절자인 PsyR에 의한 병원성 유전자들의 조절)

  • Choi, Yeon Hee;Lee, Jun Seung;Yun, Sora;Baik, Hyung Suk
    • Journal of Life Science
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    • v.25 no.2
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    • pp.136-150
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    • 2015
  • Pseudomonas syringae pathovar tabaci is a plant pathogenic bacterium that causes wildfire disease in tobacco plants. In P. syringae pv. tabaci, PsyI, a LuxI-type protein, acts as an AHL synthase, while primary and secondary sequence analysis of PsyR has revealed that it is a homolog of the LuxR-type transcriptional regulator that responds to AHL molecules. In this study, using phenotypic and genetic analyses in P. syringae pv. tabaci, we show the effect of PsyR protein as a quorum-sensing (QS) transcriptional regulator. Regulatory effects of PsyR on swarming motility and production of siderophores, tabtoxin, and N-acyl homoserine lactones were examined via phenotypic assays, and confirmed by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Further qRT-PCR showed that PsyR regulates expression of these virulence genes in response to environmental signals. However, an upstream region of the gene was not bound with purified MBP-PsyR protein; rather, PsyR was only able to shift the upstream region of psyI. These results suggested that PsyR may be indirectly controlled via intermediate-regulatory systems and that auto-regulation by PsyR does not occur.

Molecular and Ecological Analyses of Microbial Community Structures in Biofilms of a Full-Scale Aerated Up-Flow Biobead Process

  • Ju, Dong-Hun;Choi, Min-Kyung;Ahn, Jae-Hyung;Kim, Mi-Hwa;Cho, Jae-Chang;Kim, Tae-Sung;Kim, Tae-San;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.253-261
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    • 2007
  • Molecular and cultivation techniques were used to characterize the bacterial communities of biobead reactor biofilms in a sewage treatment plant to which an Aerated Up-Flow Biobead process was applied. With this biobead process, the monthly average values of various chemical parameters in the effluent were generally kept under the regulation limits of the effluent quality of the sewage treatment plant during the operation period. Most probable number (MPN) analysis revealed that the population of denitrifying bacteria was abundant in the biobead #1 reactor, denitrifying and nitrifying bacteria coexisted in the biobead #2 reactor, and nitrifying bacteria prevailed over denitrifying bacteria in the biobead #3 reactor. The results of the MPN test suggested that the biobead #2 reactor was a transition zone leading to acclimated nitrifying biofilms in the biobead #3 reactor. Phylogenetic analysis of 16S rDNA sequences cloned from biofilms showed that the biobead #1 reactor, which received a high organic loading rate, had much diverse microorganisms, whereas the biobead #2 and #3 reactors were dominated by the members of Proteobacteria. DGGE analysis with the ammonia monooxygenase (amoA) gene supported the observation from the MPN test that the biofilms of September were fully developed and specialized for nitrification in the biobead reactor #3. All of the DNA sequences of the amoA DGGE bands were very similar to the sequence of the amoA gene of Nitrosomonas species, the presence of which is typical in the biological aerated filters. The results of this study showed that organic and inorganic nutrients were efficiently removed by both denitrifying microbial populations in the anaerobic tank and heterotrophic and nitrifying bacterial biofilms well-formed in the three functional biobead reactors in the Aerated Up-Flow Biobead process.

Molecular Genetic Classification of Hypsizigus marmoreus and Development of Strain-specific DNA Markers (느티만가닥버섯의 분자유전학적 분류 및 품종특이적 DNA 마커 탐색)

  • Lim, Yun-Jeong;Lee, Chang-Yun;Park, Jeong-Eun;Kim, Sang-Woo;Lee, Hyun-Sook;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.34-39
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    • 2010
  • We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

Randomly Amplified Polymorphic DNA Analyses of Pestalotiopsis theae Isolated from Sweet Persimon (재배되는 단감나무로 부터 분리한 Pestalotiopsis theae의 RAPD 기법을 이용한 유전특성의 비교분석)

  • Lee, Youn-Su;Woo, Su-Jin;Choi, Hei-Sun;Kim, Kyoung-Su;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Chang, Tae-Hyun;Lim, Tae-Heon
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.365-372
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    • 1998
  • In this study, we evaluated the genetic relationships of fourty seven Pestalotiopsis theae isolates collected from diseased sweet persimon in various places in southern part of Korea using RAPD (Randomly Amplified Polymorphic DNAs) method. As a result of the amplification, eight primers showed total of 86 bands ranging from 0.3 Kb to 3.2 Kb. Among those 86 bands, 84 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Similarities among the compared isolates ranged from below 60% to more than 95%. Most of the compared isolates showed $50{\sim}80%$ similarities. The number of isolate pairs which showed more than 80% similarity were 248. The number of isolate pairs which showed $50{\sim}80%$ similarity were 789, and the number of isolate pairs which showed below 50% similarity were 21. Isolate SP-21 (No.9) showed below 50% similarity with all the isolates compared. At 50% similarity level, all the isolates compared, except isolate SP-21 (No.9), were included in one big group. At 65% similarity level, all the isolates compared, except isolate SP-21 (No.9), were divided into three different groups. At 75% similarity level, all the isolates compared, except isolates SP-47 (No. 23) and SP-21 (No.9), were divided into six different groups.

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Black Rot of Broccoli Caused by Xanthomonas campestris pv. campestris (Xanthomonas campestris pv. campestris에 의한 브로콜리의 검은썩음병)

  • Lee Seung-Don;Lee Jung-Hee;Kim Sun-Yee;Kim Yong-Ki;Lee Yong-Hoon;Heu Sung-Gi;Ra Dong-Soo
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.134-138
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    • 2006
  • A new bacterial disease of broccoli (Brassica oleracea var. italica) was observed on field-grown plants in Pyungchang during 2003 and 2004. Seedling infections first appeared as a blackening along the margins of the cotyledon. Cotyledon shriveled and dropped off. Infected seedlings were stunted and yellowed and eventually died. The disease was easily recognized by the presence of yellow, V-shaped, or U-shaped areas extending inward from margin of the leaf. As the disease progressed, the yellow lesions turned brown and the tissues died. Isolations made from diseased leaves on yeast extract dextrose calcium carbonate agar yielded nearly pure cultures of a yellow-pigmented bacterium typical of a xanthomonad. Two bacterial strains were purified and used for further tests. Pathogenicity of strains was confirmed on 3-week-old crucifer (cabbage, Chinese cabbage, kale, radish and broccoli) plants cut by scissors with bacterial suspensions containing $10^8 cfu/ml$ of phosphate buffered saline. The Biolog and fatty acid analyses and 16S rDNA sequencing of two strains (SL4797 and SL4800) from broccoli black rot showed that they could be identified as X. campestris pv. campestris because of their high similarity to the tester strain (X. campestris pv. campestris NCPPB528) with a match probability of 100%. This is the first report of black rot of broccoli in Korea.

Breeding of Phalaenopsis 'SM 333' with Mini Multiple Flower Formation (소형 다화 분지성 호접란 'SM 333' 육성)

  • Park, No Eun;Son, Beung Gu;Kim, Hong Yul;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.149-154
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    • 2015
  • A new Phalaenopsis cultivar 'SM 333' was bred by Sangmiwon Orchid, Korea, which produces young plants through tissue culture techniques. The new cultivar 'SM 333', showing the phenotype of multiflora with pink color and small, multibranching-type characteristics, was derived from crossing between Phalaenopsis 'Odoriko' and 'Be Tris'. An elite individual number '02-03-33' later termed 'SM 333' was selected among about 300 individual progenies, based on an intensive selection process covering vegetative and flowering distinctiveness over more than 2 years. In year 2004-2005, the 1st and 2nd characteristic analyses were carried out through performance and uniformity tests. 'SM 333' shows flower color that is bright clean pink (RHS # RP69D) and flower shape that is formal type with 5.0 and 5.8 cm in flower height and width, respectively. 'SM 333' is regarded as raceme flower type suitable for the small casual flower market. The leaves of 'SM 333' grow horizontally and about 20.8 cm in length and 6.5 cm in width. This cultivar also possesses no genetic variation, and is amenable to fast in vitro propagation and easy growth due to its vigorous growth habit. This 'SM333' was registered (Reg. # 2916) with Korea Seed & Variety Service (KSVS) on 1st December, 2009, and the plant breeder's right is currently controlled by Sangmiwon Orchid Company, Korea.

A Study on the Natural Interspecific Hybrid between Rhodeus notatus and R. ocellatus (떡납줄갱이 Rhodeus notatus와 흰줄납줄개 R. ocellatus의 자연 종간잡종에 관한 연구)

  • Yun, Bong Han;Sung, Mu Sung;Kim, Yong Hwi;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.33 no.3
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    • pp.157-166
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    • 2021
  • Three specimens, presumed to be natural hybrids between Rhodeus notatus and R. ocellatus, were collected from Onyangcheon Stream area, Jwabu-dong, Asan-si, Chungcheongnamdo, the Republic of Korea, and morphological and phylogenetic analyses were performed to clearly identify their parent species. The body color of the three natural hybrids was light greenish-brown on dorsal side, and the size of red area on the upper front of the dorsal fin and the outer margin of the anal fin generally showed intermediate characters between the parent species, R. notatus and R. ocellatus. Among the measurement and meristic characters, the ratio of prepectoral length and preanal length in the standard length, and the ratio of snout length, interorbital width, length of caudal peduncle and depth of caudal peduncle in the head length, and the number of longitudinal row scales were analyzed as the unique characters of natural hybrids. In the rag1 gene of nuclear DNA, the three natural hybrids were analyzed to be reflected all the single nucleotide polymorphism sites between R. notatus and R. ocellatus, and in the phylogenetic tree using the cytb gene of mitochondrial DNA, they formed the same genetic clade as R. notatus. Therefore, three specimens, presumed to be natural hybrids analyzed in this study were identified as interspecific hybrids between female R. notatus and male R. ocellatus.