• Title/Summary/Keyword: Gene characterization

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Characterization of Sources of Resistance to Bacterial Spot in Capsicum Peppers (고추 세균성점무늬병 저항성 유전자원과 그 주요 특성)

  • Byeon, Si-Eun;Abebe, Alebel Mekuriaw;Jegal, Yoon-Hyuk;Wai, Khin Pa Pa;Siddique, Muhammad Irfan;Mo, Hwang-Sung;Yoo, Hee Ju;Jang, Kil-Su;Hwang, Ji-Eun;Jeon, Su-Gyeong;Lee, Su-Heon;Kim, Byung-Soo
    • Horticultural Science & Technology
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    • v.34 no.5
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    • pp.779-789
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    • 2016
  • A total of 33 accessions of pepper (Capsicum spp.), including previously reported and newly discovered sources of resistance to bacterial spot caused by Xanthomonas euvesicatoria, were evaluated for their resistance to bacterial spot. The selected accessions were then grown and their horticultural characteristics were recorded. In a test for hypersensitive resistance (HR) to four races (P1, P3, P7, P8) of the pathogen found in Korea, KC00939 and Chilbok No.2, which carry the Bs2 gene, exhibited a hypersensitive response to all four races, as expected. Chilbok No.3, which carries the Bs3 gene, showed a hypersensitive reaction to race 1 and 7, as expected. KC00939 exhibited a high ASTA color value and tolerance to multiple infections from a viral complex of Cucumber mosaic virus (CMV) and Broad bean wilt virus (BBWV). Thus, this accession represents a promising genetic resource for breeding cultivars with multiple disease resistance and strong red coloration. KC01327, KC01617, KC01015, KC01760, KC01779, KC01137, KC01328, KC01006, KC00127, KC01704, and KC00995 did not exhibit hypersensitivity but showed a high level of general resistance when evaluated by spray inoculation. KC01617, KC01760, KC01779, KC01137, KC01704, and KC01777 are newly identified sources of resistance to bacterial spot. The previously and newly identified sources of resistance to bacterial spot evaluated in this study, including information about their resistance to CMV and BBWV complex in the field, the contents of pungent and sweet taste components, and the color values of dry fruits, will be useful for breeding pepper cultivars with resistance to bacterial spot.

Isolation and Characterization of Bacillus Species Having Antifungal Activity Against Pathogens of Ginseng Damping Off (인삼모잘록병원균에 항균활성을 갖는 Bacillus 균의 분리 및 특성조사)

  • Park, Kyeong Hun;Park, Hong Woo;Lee, Seong Woo;Lee, Seung Ho;Myung, Kyung Sun;Lee, Sang Yeob;Song, Jaekyeong;Kim, Young Tak;Park, Kyoung Soo;Kim, Young Ock
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.380-387
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    • 2016
  • This study was performed to select potentially available biological control agent from soil bacteria for prevention of ginseng damping off. More than five hundred strains were isolated from ginseng rhizosphere soil. By testing antifungal activity, we have selected three soil bacteria strains and their ability to produce antibiotics and lytic enzymes such as cellulase, protease and pectate lyase was examined. Also, the presence of genes for biosynthesis of lipopeptide such as fengycin, bacillomycin D, surfactin, iturin A, and zwittermicin A was investigated in selected strains. All three strains produced cellulase, protease, and xylanase. Moreover, these strains had gene for biosynthesis of bacillomycin D, surfactin, and iturin A. ES1 and ES3 strains were identified Bacillus methylotrophucus and ES2 was confirmed Bacillus amyloliquefaciens using phylogenetic analysis on the basis of 16S rRNA gene sequences. In field test, control value of ES1, ES2 and ES3 treatment was 32.4%, 46.8% and 36.7%, respectively. This results indicate that antagonistic microbes with high ability of antifungal and lytic enzyme activity can be used as a useful biological control agent to control ginseng damping off.

Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Analysis of phenotypic characterization of segregation population developed by crossing in Flammulina velutipes (팽이버섯(Flammulina velutipes) 교잡분리집단의 형태학적 특성 분석)

  • Woo, Sung-I;Kong, Won-Sik;Kim, Eun-Seou;Jang, Kab yeul;Shin, Pyung-Gyun;Oh, Youn-Lee;Oh, Min ji;Nam, Youn-keol;Kim, Kyung-Soo
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.217-222
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    • 2015
  • This study was conducted to obtain a growth correlation of basal information from the development of disease resistant Flammulina velutipes cultivars through back-crossing between the strains of wild-type brown monokaryon 4019-20 and the derivative of commercial quality white monokaryons 3. The two strains were selected to back-cross for further enhancing their latent attributes and growth characteristics. The parents of 4019-$20{\times}M3$ back-crossed to reproduce $F_1$, M3-Sn. Using $F_1$, M3-Sn procured and isolated into 94 monokaryon strains. Further examination of growth characteristics carried out by back-crossing between M3 and $BC_1F_1$ from M3-n dikaryon. Monokaryon exhibited an irregular growth pattern and demonstrated to be sluggish development in the sawdust medium. However $BC_1F_1$(M3-n) dikaryon strains confirmed mostly regular growth pattern and demonstrated ordinary growth in the sawdust medium. The fruitbody of $BC_1F_1$ confirmed as light-brown colour to be the dominant gene. The colour distributions of fruitbody, $BC_1F_1$, resulted as follows; 7% of dark brown, 25% of brown, 27% of light brown, 16% of ivory and 25% of white. The ratio of the other color to white showed 3 to 1 which suggested two major genes were related to fruitbody color.

Characterization of Bacillus thuringiensis subsp. aizawai CAB109 isolate with bioactivities to Spodoptera litura and Spodoptera exigua (Lepidoptera: Noctuidae) (담배거세미나방과 파밤나방에 활성이 있는 Bacillus thuringiensis subsp. aizawai CAB109 균주의 특성)

  • Kim, Tae-Hwan;Kim, Da-A;Kim, Ki-Su;Seo, Mi-Ja;Youn, Young-Nam;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.48 no.4
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    • pp.509-517
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    • 2009
  • Bacillus thuringiensis subsp. aizawai CAB109 isolated in Korea is known active against Spodoptera sp.. Especially, B. thuringiensis aizawai CAB109 isolates showed 100% mortality against Spodoptera litura and Spodoptera exigua. To screen highly active B. thuringiensis, the pathogenicity of B. thuringiensis CAB109 was compared with that of commercialized B. thuringiensis products. $LC_{50}$ values of CAB109, product TB-WP and product SC strains of B. thuringiensis were $1.3{\times}10^5$, $2.3{\times}10^6$ and $5.2{\times}10^5\;cfu/ml$ against the 2nd larva of S. litura and $1.8{\times}10^4$, $1.3{\times}10^6$ and $1.5{\times}10^6\;cfu/ml$ against the 2nd larva S. exigua, respectively. To determine new gene's existence and absence, the plasmid DNA was extracted, and compared to that of B.t. aizawai HD-133. Both B. thuringiensis were not like plasmid DNA pattern. PCR technique was used to predict both plasmid DNA's cry gene. PCR products analysis showed that B.t. CAB109 harbor Cry1Aa, Cry1Ab, Cry1C and Cry1D and B.t. HD-133 has Cry1Aa and Cry1Ab, respectively.

Isolation and Characterization of vasa Gene of Triploid and Diploid Human Lung Flukes (Paragonimus westermani) (폐흡충의 이배체와 삼배체 vasa 유전자 분석 및 특징)

  • Lee, Keun-Hee;Yu, Hak-Sun;Hur, Jae-Won;Yu, Sung-Suk;Choi, Sun-Hee;Park, Sang-Kyun;Lee, Sun-Joo;Chung, Dong-Il;Kong, Hyun-Hee;Ock, Mee-Sun;Jeong, Hae-Jin
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.462-469
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    • 2007
  • In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.

Characterization of Exolytic GH50A β-Agarase and GH117A α-NABH Involved in Agarose Saccharification of Cellvibrio sp. KY-GH-1 and Possible Application to Mass Production of NA2 and L-AHG (Cellvibrio sp. KY-GH-1의 아가로오스 당화 관련 엑소형 GH50A β-아가레이즈와 GH117A α-NABH의 특성 및 NA2와 L-AHG 양산에의 적용 가능성)

  • Jang, Won Young;Lee, Hee Kyoung;Kim, Young Ho
    • Journal of Life Science
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    • v.31 no.3
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    • pp.356-365
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    • 2021
  • Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

Characterization of Dopamine Receptor D4 Gene Polymorphisms in Horses (말에서 Dopamine Receptor D4 유전자의 변이 특성 분석)

  • Choi, Jae-Young;Choi, Yeonju;Lee, Jongan;Shin, Sang-Min;Yoon, Minjung;Kang, Yong-Jun;Shin, Moon-Cheol;Yoo, Ji-Hyun;Kim, Hyeonah;Cho, In-Cheol;Yang, Byoung-Chul;Kim, Nam-Young
    • Journal of Life Science
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    • v.32 no.1
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    • pp.29-35
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    • 2022
  • This study was conducted to analyze the genetic polymorphisms of dopamine receptor D4 (DRD4) in horse breeds and its association with substrate characteristics in Jeju crossbreds (Jeju Horse × Thoroughbred). Polymorphisms in DRD4 are candidate genes associated with temperament in various mammals, including humans. Single nucleotide polymorphism (SNP) G292A in the exon 3 region of the horse DRD4 has a reported association with curiosity and vigilance in thoroughbreds. Sanger sequencing was used to identify polymorphisms of the mutations in DRD4 in three horse breeds. The SNP frequency in Jeju horses was significantly different from the frequency in other breeds. Character evaluation, conducted in the Jeju crossbreds and scored using a temperament test and contact test, revealed a high correlation between each test. Comparison of the polymorphism in the DRD4 of horses and the results of the character evaluation revealed lower scores for all temperaments in horses carrying allele A. Comparison of the SNP of G292A and blood dopamine levels in Jeju crossbreds showed 2.87 times higher levels for the GA type than for the GG type. This study identified an association between DRD4 polymorphism and various test methods for evaluating horse temperament and levels of neurotransmitters. Further research could validate the use of this gene as a genetic marker for character evaluation.

Genetic Characterization of Antigenic Variant Infectious Bursal Disease Virus (IBDV) in Chickens in Korea

  • Jong-Yeol Park;Ki-Woong Kim;Ke Shang;Sang-Won Kim;Yu-Ri Choi;Cheng-Dong Yu;Ji-Eun Son;Gyeong-Jun Kim;Won-Bin Jeon;In-Hwan Kim;Bai Wei;Min Kang;Hyung-Kwan Jang;Se-Yeoun Cha
    • Korean Journal of Poultry Science
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    • v.50 no.4
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    • pp.231-240
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    • 2023
  • Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, and causes considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant IBDV (avIBDV) was reported in chicken farms in the United States. Recently, a novel avIBDV exhibited clear differences in molecular characteristics compared with previous variant strains. This study investigated the molecular characteristics of recently isolated avIBDV strains in Korea. Strains of avIBDV were confirmed by reverse transcription PCR (RT-PCR) and were propagated in 10-day-old specific-pathogen-free (SPF) embryonated chicken eggs through chorioallantoic membrane (CAM) inoculation. Multiple sequence alignment and phylogenetic analyses of hypervariable regions VP2 gene revealed that the strains originated from two different avIBDV lineages (G2a and G2d). In our results, we confirmed the co-existence and prevalence of avIBDV genogroup G2a and G2d in chicken farms. It is necessary to study the protective efficacy of current vaccines against avIBDVs.

Characterization of Traits Related to Grain Shape in Korean Rice Varieties (국내 육성 벼 품종 입형 관련 특성 분석)

  • Lee, Chang-Min;Lee, Keon-Mi;Baek, Man-Kee;Kim, Woo-Jae;Suh, Jung-Pil;Jeong, Oh-Young;Cho, Young-Chan;Park, Hyun-Su;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.199-213
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    • 2020
  • Grain size and shape are the two important components contributing to rice yield and quality. To analyze traits related to grain-shape, a total of 272 varieties derived from japonica, japonica black and Tongil-type rice accession in Korea were evaluated in this study. The traits, grain length (GL), grain width (GW), grain thickness (GT), length to width ratio (RLW), and 1000-grain weight (TGW) were measured and replicated 10 times. Genes (GW2, GS3, qGL3, qSW5, GS5, TGW6, GW7, and GW8) related to grain-shape were validated in the accessions using specific DNA marker sets. K-mean clustering of the accession based on phenotypic data revealed three groups: group 1 was classified by GW and GT and included most of japonica type, group 2 was classified by RLW and GL reached a medium size and possessed a half spindle-shaped type, and group 3 was classified by TGW, reached a long size and possessed a semi-round shape. In validation tests using the marker sets, both gw2 and tgw6 were validated in less than 1% of the tested accessions and two allelic types, qgl3 and gw8, were only verified in Tongil-type accessions. For GW8 and GW2, any different amplicons were not amplified in any japonica or Tongil-type accessions, respectively. In order to suggest the representative grain-shape gene combinations for each ecotype, the allelic combinations were evaluated by PCR analysis. Cj1 and 2 in japonica (Cj1-7), Cj_b1 and 2 in japonica-black (Cj_b1-3), and CT3 in Tongil-type (CT1-13) turned out to be the dominant combination in each ecotype, respectively. In addition, the results revealed that introgression of four genes (gw2, gs3, qSW5, and GS5) would expand the diversity of grain shape in Korean japonica varieties. The gene combinations information could be utilized practically to understand or enhance grain shape in japonica rice breeding program.