• 제목/요약/키워드: rDNA PCR-RFLP

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Phytophthora nicotianae에 의한 국내 미기록 화훼류 역병 (Unrecorded Phytophthora Diseases of Flowering Plants Caused by Phytophthora nicotianae in Korea)

  • 지형진;김완규;김재영;임성언
    • 한국식물병리학회지
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    • 제14권5호
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    • pp.452-457
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    • 1998
  • Thirty-eight isolates of Phytophthora sp. caused rots on roots and basal stems were collected from five flowering plants from 1992 to 1997 at eight cultivation areas in Korea. All the isolates were identified as P. nicotianae based on following characteristics. The fungus produced markedly papillate, not caducous and ovoid to spherical sporangia, abundant chlamydospores, and small oospores with amphigynous antheridia only when paired with either A1 or A2 mating type. All isolates grew well at 35$^{\circ}C$ and showed distinct arachnoid colony patterns on CMA and PDA. Sizes of sporangia and chlamydospores of five representative isolates from each plant averaged 43-52$\times$30-38 ${\mu}{\textrm}{m}$ and 28 ~34 ${\mu}{\textrm}{m}$. Mating type of the isolates was either A1 or A2, and oogonia and oospores were measured as 28~31 ${\mu}{\textrm}{m}$ and 21~25 ${\mu}{\textrm}{m}$. PCR-RFLP analysis of rDNA of the five isolates resulted that restriction band patterns of the small subunit and ITS regions were identical to a perilla isolate of P. nicotianae, but distinct from P. cactorum and P. capsici. Cross inoculation tests showed that the five isolates had pathogenicity to lily, christmas cactus, anthurium, baby's breath and carnation with different degrees. However, each isolate showed stronger pathogenicity to its corresponding original host than others. Among five lily cultivars Georgia and Quririna were more susceptible than Napoli and others. This is first report of Phytophthora root and stem rot of lily, Christmas cactus, anthurium, baby's breath and monochoria in Korea.

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Identification of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Golpayegani Cattle by DNA Test

  • Mosafer, J.;Nassiry, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권12호
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    • pp.1691-1695
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    • 2005
  • The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

DNA Polymorphisms of κ-Casein, β-Lactoglobulin, Growth Hormone and Prolactin Genes in Korean Cattle

  • Chung, E.R.;Kim, W.T.;Lee, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제11권4호
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    • pp.422-427
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    • 1998
  • The gene and genotypic frequencies of ${\kappa}$-casein (${\kappa}$-CN), ${\beta}$-lactoglobulin (${\beta}$-LG), growth hormone (bGH) and prolactin (bPRL) loci in Korean cattle were investigated using PCR-RFLP analyses. Genomic DNA samples were obtained from 290 cows and 30 AI bulls. In both cows and bulls, the most predominant genotypes of ${\kappa}$-CN, ${\beta}$-LG, bGH and bPRL loci were AB, BB, AA and AA, respecitively. The frequencies of A and B alleles for ${\kappa}$-CN locus were .612 and .388 for cows and .567 and .433 for bulls. The respective frequencies of A and B alleles for ${\beta}$-LG locus were .153 and .847 in cows and .217 and .783 in bulls. The frequencies of A and B alleles for bGH locus were .769 and .231 in cows and .784 and .216 in bulls, respectively. The frequencies of A and B alleles for bPRL locus were .678 and .322 for cows and .767 and .233 for bulls. Differences in frequencies of these alleles were not significant between cows and bulls at all loci examined. If the DNA polymorphisms of these candidate genes are associated with economically important traits, they could serve as genetic markers for genetic improvement in future marker-assisted selection programs in Korean cattle.

Molecular Phylogenetic Diversity and Spatial Distribution of Bacterial Communities in Cooling Stage during Swine Manure Composting

  • Guo, Yan;Zhang, Jinliang;Yan, Yongfeng;Wu, Jian;Zhu, Nengwu;Deng, Changyan
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권6호
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    • pp.888-895
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    • 2015
  • Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and subsequent sub-cloning and sequencing were used in this study to analyze the molecular phylogenetic diversity and spatial distribution of bacterial communities in different spatial locations during the cooling stage of composted swine manure. Total microbial DNA was extracted, and bacterial near full-length 16S rRNA genes were subsequently amplified, cloned, RFLP-screened, and sequenced. A total of 420 positive clones were classified by RFLP and near-full-length 16S rDNA sequences. Approximately 48 operational taxonomic units (OTUs) were found among 139 positive clones from the superstratum sample; 26 among 149 were from the middle-level sample and 35 among 132 were from the substrate sample. Thermobifida fusca was common in the superstratum layer of the pile. Some Bacillus spp. were remarkable in the middle-level layer, and Clostridium sp. was dominant in the substrate layer. Among 109 OTUs, 99 displayed homology with those in the GenBank database. Ten OTUs were not closely related to any known species. The superstratum sample had the highest microbial diversity, and different and distinct bacterial communities were detected in the three different layers. This study demonstrated the spatial characteristics of the microbial community distribution in the cooling stage of swine manure compost.

한우 Exostosin-1 유전자의 SNP 탐색 및 경제형질 관련성 분석 (Association Study Between the Polymorphisms of Exostosin-1 Gene and Economic Traits in Hanwoo)

  • 김범수;김남국;이승환;조용민;허강녕;박응우;양부근;윤두학
    • Journal of Animal Science and Technology
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    • 제53권1호
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    • pp.7-13
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    • 2011
  • 본 연구는 한우의 경제형질 관련 유전적 표지인자(DNA marker) 개발을 목적으로 EXT1 유전자의 다형성과 발현양상을 관찰하여 경제형질과의 관련성을 확인하고자 수행하였다. PCRdirect sequencing을 통하여 4개(T272196A, C272359T, G290964A 및 A302092G)의 SNPs를 탐색 하였으며, 탐색된 SNPs를 통하여 PCR-RFLP 기법으로 유전자형을 결정한 후 경제형질과 관련성을 분석하였다. 그 결과 T272196A 좌위에서 근내지방 육종가(p=0.014), G290964A 좌위에서 등지방두께 육종가 추정치(p=0.001), A302092G 좌위에서는 등지방두께 육종가(p < 0.001) 및 등심단면적 육종가(p=0.020)에서 각각 유의적인 연관성이 인정되었다. 따라서, 본 연구를 통해 확인된 SNP를 이용하여 한우의 선발에 활용 가능할 것으로 사료된다.

Rett 증후군 34례의 MECP2 유전자 변이에 관한 연구 (Mutational Analysis of MECP2 Gene in 34 Rett Syndrome)

  • 박상조;황태규;손병희;김철민
    • Clinical and Experimental Pediatrics
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    • 제45권10호
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    • pp.1263-1272
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    • 2002
  • 목 적 : 레트증후군이란 1966년 Andreas Rett에 의해 처음 기술되었으며 생후 6개월에서 18개월 정도까지 비교적 정상 발달을 한 후 두위 발달의 감소와 함께 습득했던 인지 및 운동능력의 상실, 언어기능의 상실, 그리고 특징적인 손동작(상동행동)을 보이는 X 염색체 우성유전으로 생각되는 질환이다. 1999년 미국에서 그 결함 유전자인 MECP2 유전자가 Xq28에서 밝혀졌으나 우리 나라에서는 이 질환에 대한 전반적 이해가 부족하며 유전자 연구는 거의 전무한 상태이다. 이에 저자들은 우리 나라의 Rett 증후군 환아에서 MECP2 유전자의 결함이 어느 정도 나타나는지를 알고자 이 연구를 시행하였다. 방 법 : 2000년 2월부터 2001년 3월까지 본원을 방문한 Rett 증후군 환아 34례의 말초혈액을 EDTA (Ethylenediaminetetraacetic acid) 처리하여 5 cc 채취하여 냉동 보관한 후 해빙하여 DNA 표본을 추출했다. DNA의 추출은 E.Z.N.A blood DNA kit을 사용하였다. MECP2 유전자 네 종류의 axon은 PCR을 이용해 증폭하였고 primer sequence는 1999년 Amir 등에 의해 디자인 된 것(AF030876)을 사용하였다. ABI 377 DNA sequencer와 ABI PRISM dye cycle sequencing reaction kit을 사용하여 DNA sequencing을 시행하였고 우리 나라에서 최초로 발견된 유전자 변이에 대해서는 RFLP 분석을 통해 확인하였다. 결 과 : 1) MECP2 유전자의 변이를 보인 환아는 23례로 67.6%에서 관찰되었다. 2) 9종의 missense mutation과 3종의 nonsense mutation을 합해 총 12종의 유전자 변이가 관찰되었다. 3) 이들 돌연 변이 중 L100V, G161E, 그리고 T311M은 전세계적으로 우리 나라에서 처음 발견된 변이이다. 4) 23례의 유전자 변이는 대부분(78.3%) MBD와 TRD라는 기능영역에서 발견되었다. 5) 본 연구에서 T158M, R270X, 그리고 R306C가 자주 나타나는 유전자 변이였다. 결 론 : 우리 나라의 Rett 증후군 환아에서도 MECP2 유전자의 변이는 비교적 흔히 관찰되어 MECP2 유전자의 이상이 Rett 증후군을 유발하는 주 유전자 이상임을 확인하였고 Rett 증후군 환아의 확진을 위해 MECP2 유전자 검사가 필요할 것으로 사료된다.

Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Jong-Hyun;Kim, Yong-Hwan
    • 대한수의학회지
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    • 제54권2호
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    • pp.91-99
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    • 2014
  • An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

DNA Polymorphism of Insulin-like Growth Factor-binding Protein-3 Gene and Its Association with Cashmere Traits in Cashmere Goats

  • Liu, Haiying;Liu, Chao;Yang, Guiqin;Li, Hui;Dai, Jin;Cong, Yuyan;Li, Xuejian
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권11호
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    • pp.1515-1520
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    • 2012
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is important for regulation of growth and development in mammals. The present investigation was carried out to study DNA polymorphism by PCR-RFLP of IGFBP-3 gene and its effect on fibre traits of Chinese Inner Mongolian cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Four hundred and forty-four animals were used to detect polymorphisms in the hircine IGFBP-3 gene. A 316-bp fragment of the IGFBP-3 gene in exon 2 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in the populations. The frequency of AA, AB and BB genotypes was 0.58, 0.33 and 0.09 respectively. The allelic frequency of the A and B allele was 0.75 and 0.25 respectively. Nucleotide sequencing revealed a C>G transition in the exon 2 region of the IGFBP-3 gene resulting in R158G change which caused the polymorphism. Least squares analysis revealed a significant effect of genotypes on cashmere weight (p<0.0001), cashmere fibre length (p<0.001) and hair length (p<0.05) of the animals. The effect of genotypes on cashmere fibre diameter was not statistically significant (p>0.05). The animals of AB and BB genotypes showed higher cashmere weight, cashmere fibre length and hair length than the animals possessing AA genotype. These results suggested that polymorphisms in the hircine IGFBP-3 gene might be a potential molecular marker for cashmere weight in cashmere goats.

Stem Rot on Adzuki Bean (Vigna angularis) Caused by Rhizoctonia solani AG 4 HGI in China

  • Sun, Suli;Xia, Changjian;Zhang, Jiqing;Duan, Canxing;Wang, Xiaoming;Wu, Xiaofei;Lee, Suk-Ha;Zhu, Zhendong
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.67-71
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    • 2015
  • During late August and early September 2011, stem rot symptoms were observed on adzuki bean plants (Vigna angularis) growing in fields located in Beijing and Hebei Province, China, respectively. In this study, four isolates were obtained from infected stems of adzuki bean plants. Based on their morphology, and sequence and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses of the ribosomal DNA internal transcribed spacers (rDNA-ITS) region, the four isolates were identified as Rhizoctonia solani in anastomosis group (AG) 4 HGI. Pathogenicity tests showed that all isolates were strongly pathogenic to adzuki bean and resulted in serious wilt symptoms which was similar to observations in the fields. Additionally, the isolates infected several other crops and induced related rot on the roots and basal stems. To our knowledge, this is the first report of Rhizoctonia solani AG 4 HGI causing stem rot on adzuki bean.

참깨 엽화병의 발생과 파이토플라스마의 검출 (Occurrence of Sesame Phyllody Disease in Korea and Detection of Its Phytoplasma)

  • 한무석;노은운;윤정구
    • 한국식물병리학회지
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    • 제13권4호
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    • pp.239-243
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    • 1997
  • 1990년 8월 충북 보은의 참깨 밭에서 전형적인 엽화병(葉化病; phyllody disease)이 발생하였다. 병징은 꽃이 잎처럼 녹색으로 변하고, 이병엽은 정상엽보다 작고 잔가지가 마치 빗자루 모양으로 총생하였고 개화 및 결실이 되지 않았다. 투과 전자현미경으로 병든 잎의 사관요소에서 phytoplasma가 존재하는 것을 확인하였다. 병든 잔가지에서 추출한 DNA를 중합효소 연쇄반응으로 분석한 결과 대추나무 빗자루병에 걸린 나무에서 증폭되는 DNA와 같은 크기의 band가 관찰되었다. 따라서 위의 시료는 phytoplasma 벙으로 확인 되었으며 참깨 엽화병으로 명명하였다. 또한 참깨 엽화병을 일으키는 phytoplasma와 대추 나무 빗자루병 phytoplasma를 PCR 증폭 및 제한효소로 절단하여 분석한 결과 이들은 서로 다른 그룹이라는 것을 알 수 있었다.

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