• 제목/요약/키워드: PCR-RFLP of rDNA.

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Septobasidium sp.에 의한 구실잣밤나무 고약병의 분자학적 다양성 분석 (Analysis of Molecular Diversity in Castanopsis sieboldii with Felt Disease Caused by Septobasidium sp.)

  • 이건우;서상태;차병진;한상섭
    • 식물병연구
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    • 제29권4호
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    • pp.420-424
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    • 2023
  • 2020년 제주도 동백동산 내 구실잣밤나무 수피에 고약병과 관련된 Septobasidium sp.가 발견되었다. 분리한 균주는 습실 처리하여 새로 생성된 균사에 대한 genomic DNA를 추출한 뒤 internal transcribed spacer 및 small subunit rDNA 유전자에 대해 염기서열을 밝혔으며 polymerase chain reaction-based restriction fragment length polymorphism 분석을 통해 균주들간의 분자학적 특성이 조사되었다. 이 새로운 Septobasidium sp.은 기존에 알려진 고약병들과 형태학적 및 계통학적으로 다르게 나타나 새로운 종으로 보고한다. 또한 이 고약병은 관찰 당시 주변의 다른 수목들에서는 발생하지 않고 오직 구실잣밤 나무에서만 나타나는 특성으로 기주특이성이 매우 높다는 것이 확인되었다.

Morphometric and Genetic Variability Among Tylenchulus semipenetrans Populations from Citrus Growing Area in Korea

  • Park, Byeong-Yong;Park, Sun-Nam;Lee, Jae-Kook;Bae, Chang-Hwan
    • The Plant Pathology Journal
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    • 제25권3호
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    • pp.236-240
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    • 2009
  • Tylenchulus semipenetrans, citrus nematode is an important phytopathogenic nematode and responsible for serious damage on citrus. However, little information is available about genetic variability of T. semipenetrans among different populations with variation of conventional diagnostic characteristics. In this study, we compared the morphometric and genetic characteristics among different populations. The mature female of T. semipenetrans collected in this study had thicker cuticle than those in the previous studies. In comparative sequence analysis of T. semipenetrans populations obtained from Jeju in Korea, we observed genetic variations within clones generated from single individuals. To determine whether variability among copies of nuclear ribosomal DNA sequences exists in the genome of T. semipenetrans, PCR-RFLP technique from individuals of Korean isolates with MseI and MspI restriction enzymes was used to prove experimentally that all populations have intra-specific variations. Restriction enzyme digestion created several fragments on 3.0% agarose gel corresponding to several haplotypes in all populations, though some populations displayed fragment deletion. The total length of fragments was larger than before digestion, indicating sequence heterogeneity within the genome of T. semipenetrans.

심비디움에 발생하는 해충의 종류 (Pests occurring on Cymbidium)

  • 조명래;전성욱;강택준;김형환;안승준;양창열
    • 한국응용곤충학회지
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    • 제52권4호
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    • pp.403-408
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    • 2013
  • 수출용 심비디움에 발생하는 해충의 종류와 해충관리 현황을 알아보고자 국내 주요 재배단지 10개 지역 45농가를 대상으로 조사를 실시하였다. 농가에서 채집된 해충의 종류는 응애류(20농가), 깍지벌레류(6농가), 민달팽이류(6농가), 총채벌레류(8농가), 진딧물류(5농가) 노린재류(1농가) 등이었다. 채집한 해충은 현미경을 이용하여 동정한 결과 점박이응애(Tetranychus urticae Koch), 대만총채벌레(Frankliniella intonsa Trybom), 난초핀깍지벌레(Pinnaspis aspidistrae Signoret), 민달팽이(Incilaria confusa Cockarel), 썩덩나무노린재(Halyomorpha brevis Walker), 복숭아혹진딧물(Myzus persicae Sulzer), 목화진딧물(Aphis gossypii Glover), 무화과깍지벌레(Coccus hesperidum Linnaeus)와 소형달팽이류 1종으로 나타났다. 총채벌레 산란피해로 의심되는 증상의 정확한 동정을 위해 rDNA의 ITS2 부위를 PCR-RFLP 한 결과 아까시총채벌레(Thrips flavus Schrank)와 파총채벌레(Thrips tabaci Lindeman)로 확인되었다. 따라서, 수출 심비디움을 재배하는 농가에서는 이러한 해충들에 대한 종합적인 방제대책을 수립하여 실천하는 것이 필요할 것으로 생각된다.

Genetic Variation and Species Identification of Thai Boesenbergia (Zingiberaceae) Analyzed by Chloroplast DNA Polymorphism

  • Techaprasan, Jiranan;Ngamriabsakul, Chatchai;Klinbunga, Sirawut;Chusacultanachai, Sudsanguan;Jenjittikul, Thaya
    • BMB Reports
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    • 제39권4호
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    • pp.361-370
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    • 2006
  • Genetic variation and molecular phylogeny of 22 taxa representing 14 extant species and 3 unidentified taxa of Boesenbergia in Thailand and four outgroup species (Cornukaempferia aurantiflora, Hedychium biflorum, Kaempferia parviflora, and Scaphochlamys rubescens) were examined by sequencing of 3 chloroplast (cp) DNA regions (matK, psbA-trnH and petA-psbJ). Low interspecific genetic divergence (0.25-1.74%) were observed in these investigated taxa. The 50% majority-rule consensus tree constructed from combined chloroplast DNA sequences allocated Boesenbergia in this study into 3 different groups. Using psbA-1F/psbA-3R primers, an insertion of 491 bp was observed in B. petiolata. Restriction analysis of the amplicon (380-410 bp) from the remaining species with Rsa I further differentiated Boesenbergia to 2 groupings; I (B. basispicata, B. longiflora, B. longipes, B. plicata, B. pulcherrima, B. tenuispicata, B. thorelii, B. xiphostachya, Boesenbergia sp.1 and Boesenbergia sp.3; phylogenetic clade A) that possesses a Rsa I restriction site and II (B. curtisii, B. regalis, B. rotunda and Boesenbergia sp.2; phylogenetic clade B and B. siamensis; phylogenetic clade C) that lacks a restriction site of Rsa I. Single nucleotide polymorphism (SNP) and indels found can be unambiguously applied to authenticate specie-origin of all investigated samples and revealed that Boesenbergia sp.1, Boesenbergia sp.2 and B. pulcherrima (Mahidol University, Kanchanaburi), B. cf. pulcherrima1 (Prachuap Khiri Khan) and B. cf. pulcherrima2 (Thong Pha Phum, Kanchanaburi) are B. plicata, B. rotunda and B. pulcherrima, respectively. In addition, molecular data also suggested that Boesenbergia sp.3 should be further differentiated from B. longiflora and regarded as a newly unidentified Boesenbergia species.

Ten Cases of Taenia saginata Infection Confirmed by Analysis of the Internal Transcribed Spacer 1 rDNA Region in the Republic of Korea

  • Song, Su-Min;Yun, Hae Soo;VanBik, Dorene;Chang, Hyun-Ha;Lee, Sang-Ah;Kim, Shin-Woo;Ryoo, Namhee;Eun, Dong Yeub;Lee, Nan Young;Goo, Youn-Kyoung;Hong, Yeonchul;Ock, Meesun;Cha, Hee-Jae;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • 제57권4호
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    • pp.417-422
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    • 2019
  • From October 2015 to August 2018, tapeworm proglottids were obtained from 10 patients who were residents of Daegu and Gyeongbuk provinces and had a history of raw beef consumption. Most of them had no overseas travel experience. The gravid proglottids obtained from the 10 cases had 15-20 lateral uterine branches. A part of internal transcribed spacer 1 (ITS1) DNA of the 10 cases, amplified by polymerase chain reaction (PCR) and digested with AleI restriction enzyme, produced the same band pattern of Taenia saginata, which differentiated from T. asiatica and T. solium. Sequences of ITS1 and cytochrome c oxidase subunit 1 (cox1) showed higher homology to T. saginata than to T. asiatica and T. solium. Collectively, these 10 cases were identified as T. saginata human infections. As taeniasis is one of the important parasitic diseases in humans, it is necessary to maintain hygienic conditions during livestock farming to avoid public health concerns.

한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성 (Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan)

  • 홍석원;문지영;이복권;김영부
    • 생명과학회지
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    • 제17권3호통권83호
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    • pp.386-395
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    • 2007
  • 본 연구는 1999년에서 2001년도 걸쳐서 3년간 국내에서 분리된 환자유래 장염비브리오 18균주와 일본 후쿠오카 지역에서 2002년도에 환자에서 분리한 장염비브리오 9균주 등 총 27균주에 대하여 toxR 유전자의 검출, 혈청형별 검사, 약제내성의 양상, tdh, trh1 및 trh2 유전자의 보유상태 및 urease 생성성을 살펴보고, 혈청형 O3:K6 균주에 대하여 TDH의 생성성 검사, tdh 양성균주의 RFLP 형별, ORF 8의 분포, PFGE법과 RAPD법을 실시하였으며 결과는 다음과 같다. 1. 한국 및 일본의 환자유래 균주 대부분에서 urease음성이었으며, toxR 유전자로 확인 동정하였고 혈청형의 분포는 국내 분리 주의 O3:K6, O4:K9, O6:K46, O3:K57, O5:Kl5와 일본 분리주의 O3:K6, O1:K38, O4:K68, O4:Kl2의 혈청형으로 나타났다. 2. 항생제 감수성 시험에서는 vancomycin과 oxacillin은 27균주 (100%), penicillin은 26균주 (96.3%)로 높은 내성을 나타내었고, 14균주 (51.9%)가 4가지 이상의 항생제에 다제내성을 나타내었다. 항생제의 내성양상은 6종류로 혈청형에 관계없이 vancomycin, oxacillin, penicillin에 내성을 나타내는 V형이 15균주 (55.6%)로 가장 많이 나타났다. 3. tdh 유전자는 26균주가 PCR법과 DNA probe hybridization법의 결과에서 양성으로 나타났으며, urease양성이었던 일본 환자유래 혈청형 O3:K6 1균주만이 tdh유전자 음성, trh2 유전자 양성을 나타냈다. 혈청형 O3:K6의 TDH 생성성역가는 전 균주가 256배에서 2,048배 정도로 나타났으며, RFLP 양상은 모든 균주가 11.5 kb와 4.3 kb에 tdh 유전자를 보유하고 있었다. 또한 혈청형 O3:K6 균주들은 RAPD법과 PFGE법으로 유전자형별을 비교 검토한 결과 8형으로 거의동일한 유전자형별의 결과를 나타내었다. 4. ORF 8의 분포는 혈청형 O3:K6 전 균주에서 양성이었고, 특히 혈청형 O6:K46 4균주 모두에서 ORF 8 양성을 나타내어 새롭게 나타난 유행균주일 가능성을 시사 하였다. 따라서 ORF 8 유전자의 검출은 범세계적으로 유행하는 균주들을 동정하는데 있어 genetic marker로서 매우 유용한 것으로 사료된다.

Association of Paraoxonase-1(Q192R and L55M) Gene Polymorphisms and Activity with Colorectal Cancer and Effect of Surgical Intervention

  • Ahmed, Nagwa S.;Shafik, Noha M.;Elraheem, Omar Abd;Abou-Elnoeman, Saad-Eldin A.
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권2호
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    • pp.803-809
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    • 2015
  • Background: Colorectal cancer (CRC) is a leading cause of cancer-related death. Oxidative DNA damage may contribute to cancer risk and the antioxidant paraoxonase is one endogenous free radical scavenger in the human body which could therefore exert an influeence. Purpose: Aim of this study was to determine the role of serum arylesterase (ARE) and paraoxonase 1(PON1) activities in CRC patients and to find any association between (PON1) Q192R and L55M gene polymorphisms in CRC patients. Also the serum ARE and PON1 activities in CRC patients will be investigated before and after surgery Materials and Methods: This study involved a total of 50 patients with newly diagnosed CRC and 80 healthy controls. PON1 and ARE activities were determined using an enzymatic spectrophotometric method. PON1 Q192R and L55M gene polymorphisms were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based restriction fragment analysis. The restriction enzyme AlwI was used to examine the Q192R polymorphism and Hsp92II for the L55M polymorphism. Results: Significant differences in the PON1 Q192R polymorphism were found between patients and controls. The Q allele was more frequent in the patient group than in controls, while the R allele was more frequent in the controls. Significant differences were found in the L55M polymorphism. Additionally, there were significant differences in L and M allele frequencies (p=0.001). The serum activities of PON1 and ARE were low in QQ and MM genotype. Conclusions: serum PON1 and ARE activities were significantly lower in CRC patients compared to healthy subjects. The R allele may protect against colorectal cancer.

Diversity of Epiphytic and Acid-tolerant Epiphytic Bacterial Communities on Plant Leaves

  • Joung Pil-Mun;Shin Kwang-Soo;Lim Jong-Soon;Park Seong Joo
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2002년도 추계학술대회
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    • pp.100-105
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    • 2002
  • The diversity of epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic deposition to the phyllosphere using 16S rDNA sequence data. In addition, acid-tolerant epiphytic bacterial communities were compared. A total of 78 epiphytic and 444 acid-tolerant clones were obtained from clone libraries, resulting in 20 and 17 phylotypes by analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A low bacterial diversity in both areas was found. As tree leaves grow older, bacterial diversities were slightly increased in the level of subphylum. The community structure of epiphytic bacteria in both areas in April consisted of only two subphyla, $\beta-and\;\gamma-Proteobacteria$. In August two additional subphyla in both areas were found, but the composition was a little different, Acidobacteria and Cytophaga-Flexibacter-Bacteroids (CFB) group in the industrial estate and a -Proteobacteria and CFB group in the natural area, respectively. Acidobacteria could be an indicator of epiphytic bacteria for acidic deposition on plant leaves, whereas a -Proteobacteria be one of epiphytic bacteria that naturally survive on leaves that are not affected by acidic deposition. The acid-tolerant bacterial communities in April were composed of two subphyla, $\gamma-Proteobacteria$ and Low G+C gram-positive bacteria in both areas, and in August a-Proteobacteria was added to the community just in the natural forest area. The direct influence of acidic deposition on the acid-tolerant bacterial phylogenetic composition could not be detected in higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group of $\gamma-Proteobacteria$ just in the industrial estate.

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Association of a miR-502-Binding Site Single Nucleotide Polymorphism in the 3'-Untranslated Region of SET8 and the TP53 Codon 72 Polymorphism with Cervical Cancer in the Chinese Population

  • Yang, Shao-Di;Cai, Yan-Lin;Jiang, Pei;Li, Wen;Tang, Jian-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권16호
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    • pp.6505-6510
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    • 2014
  • Objective: This study was conducted to identify whether polymorphic variants of set domain-containing protein 8 (SET8) and tumor protein p53 (TP53) codon 72, either independently or jointly, might be associated with increased risk for cervical cancer. Methods: We genotyped SET8 and TP53 codon 72 polymorphisms of peripheral blood DNA from 114 cervical cancer patients and 200 controls using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct DNA sequencing. Results: The frequency of SET8 CC (odds ratios (OR) = 2.717, 95% CI=1.436-5.141) or TP53 GG (OR=2.168, 95% CI=1.149-4.089) genotype was associated with an increased risk of cervical cancer on comparison with the SET8 TT or TP53 CC genotypes, respectively. In additional, interaction between the SET8 and TP53 polymorphisms increased the risk of cervical cancer in a synergistic manner, with the OR being 9.913 (95% CI=2.028-48.459) for subjects carrying both SET8 CC and TP53 GG genotypes. Conclusion: These data suggest that there are significant associations between the miR-502-binding site SNP in the 3'-UTR of SET8 and the TP53 codon 72 polymorphism with cervical cancer in Chinese, and there is a gene-gene interaction.

Screening of BCL-2 associated X protein gene polymorphism associated with scrotal hernia in domesticated swine using polymerase chain reaction-restriction fragment length polymorphism

  • Manalaysay, Jessica G.;Antonio, Nathaniel D.;Apilado, Ralph Lorenz R.;Bambico, Joseph F.;Mingala, Claro N.
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권2호
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    • pp.262-266
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    • 2017
  • Objective: This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods: Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results: Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion: Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd.