• 제목/요약/키워드: Species-specific PCR

검색결과 648건 처리시간 0.024초

Identification of Papaya Ringspot Potyvirus type W infecting squash in Korea

  • T. S. Jin;Lee, S. H.;Park, J. W.;Park, H.S.;Kim, M.;D. B. Shin;J. U. Cheon;B. J. Cha
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.141.2-142
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    • 2003
  • A flexuous rod-shaped virus was isolated from Cucurbita pepo leaves showing green mosaic and puckering symptoms at Anseong, Korea. Based on the biological tests, electron microscopy, and reverse transcription-polymerase chain reaction (RT-PCR), the isolate was identified as Papaya ringspot virus type Watermelon (PRSV-W). In the biological test, host range of PRSV-W was limited in the families Cucurbitaceae and Chenopodiaceae. Most susceptible cucurbit species, such as Cucurmis lanatus, Cucurmis sativus, Cucurbita pepo, and Citrullus lanatus, responded to mechanical inoculation by PRSV-W that induce green mosaic, malformation, puckering, and narrow laminae. The local lesion symptoms were produced on the inoculated leaves of Chenopodium maranticolor and C. quinoa PRSV specific primers which amplifies the part of the coat protein (CP) genes, generated a 648 bp product from 6 isolates of PRSV-W, but no amplification had been detected in other viruses including CMV, CGMMV, KGMMV, ZYMV and WMV. In electron microscopy, PRSV particles were flexuous, approximately 780 nm in length and 12 nm in width. PRSV-W is one of the worldwide viruses which has the great economic importance in cucumber, melon, squash, watermelon, and other cultivated cucurbits with ZYMV and WMV. This is the first report of PRSV-W on cucurbits in Korea.

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구강 편평상피암종에서 CDH-13 유전자의 promoter methylation에 대한 연구 (PROMOTER METHYLATION OF THE CDH-13 GENE IN THE ORAL SQUAMOUS CELL CARCINOMA)

  • 이문주;한세진;김경욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제34권5호
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    • pp.525-531
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    • 2008
  • CDH-13(T-cadherin), which is one of a kind among the 20 cadherins, can be found mainly in wall of aorta, neuron, spleen, blood vessel etc. It is also called H-cadherin. This structural difference can explain that CDH-13 is thought to play a key role in maintaining mutual relation between extra and intra-cellular environment rather than in cell adhesion. The main function of CDH-13 is to participate in blood vessel function. Additionally, it is known to regulate cell growth and cell contact inhibition. When cells are proliferating, cell surface perceives other cells so that substance such as CDH-13 can inhibit their growth or proliferation resulting in homeostasis without endless proliferation or invasion of connective tissue boundaries. However, tumor cell itself appears to be different from normal cells' growth, invasion or transmission. Therefore, it can be diagnosed that these characteristics are closely related to expression of CDH-13 in tumor cells. This study is to investigate expression of CDH-13 in SCC and its correlation with promoter methylation. 20 of tissue species for the study are excised and gathered from 20 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find development of CDH-13 in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1.Immunohistochemical staining: In normal oral squamous epithelial tissue, strong expression of CDH-13 was found in cell plasma membrane of basal cell layer. On the other hand, in case of low-differentiated oral SCC, development of CDH-13 was hardly seen. 2.The development of CDH-13 gene: In 9 of samples, expression of CDH-13 gene could be seen and 2 of them showed low expression compared to the others. And rest of the 11 samples showed no expression of CDH-13 gene. 3.Methylation of CDH-13 gene: Among 9 samples which expressed CDH-13 gene, 7 of them showed unmethylation. In addition, among 11 samples without CDH-13 gene expression, 10 showed methylation. According to the results stated above, promoter methylation were found in 13 samples(65%) among 20 of oral SCC samples. In low-differentiated SCC, suppression of gene expression could be seen accompanying promoter methylation. These phenomenon of gene expression was proved by immunohistochemical investigation. Finally, for development of oral SCC, conclusions can be made that suppression of CDH-13 played a main role and suppression of gene expression was originated from promoter methylation. Considering this, it is expected that suppression of CDH-13 from promoter methylation to be utilized as a good diagnostic marker of oral SCC.

Modulated Gene Expression of Toxoplasma gondii Infected Retinal Pigment Epithelial Cell Line (ARPE-19) via PI3K/Akt or mTOR Signal Pathway

  • Zhou, Wei;Quan, Juan-Hua;Gao, Fei-Fei;Ismail, Hassan Ahmed Hassan Ahmed;Lee, Young-Ha;Cha, Guang-Ho
    • Parasites, Hosts and Diseases
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    • 제56권2호
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    • pp.135-145
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    • 2018
  • Due to the critical location and physiological activities of the retinal pigment epithelial (RPE) cell, it is constantly subjected to contact with various infectious agents and inflammatory mediators. However, little is known about the signaling events in RPE involved in Toxoplasma gondii infection and development. The aim of the study is to screen the host mRNA transcriptional change of 3 inflammation-related gene categories, PI3K/Akt pathway regulatory components, blood vessel development factors and ROS regulators, to prove that PI3K/Akt or mTOR signaling pathway play an essential role in regulating the selected inflammation-related genes. The selected genes include PH domain and leucine- rich-repeat protein phosphatases (PHLPP), casein kinase2 (CK2), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), glutamate-cysteine ligase (GCL), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase (NQO1). Using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), we found that T. gondii up-regulates PHLPP2, $CK2{\beta}$, VEGF, GCL, GST and NQO1 gene expression levels, but down-regulates PHLPP1 and PEDF mRNA transcription levels. PI3K inhibition and mTOR inhibition by specific inhibitors showed that most of these host gene expression patterns were due to activation of PI3K/Akt or mTOR pathways with some exceptional cases. Taken together, our results reveal a new molecular mechanism of these gene expression change dependent on PI3K/Akt or mTOR pathways and highlight more systematical insight of how an intracellular T. gondii can manipulate host genes to avoid host defense.

광주지역 도축 돼지 및 가공품 E형 간염 실태 조사 (Screening of slaughter pig and pork products for hepatitis E virus in Gwangju and nearby areas)

  • 정하진;김지연;최인수;성창민;박자윤;박지영;안아진;곽진주;장미선;서계원;김용환
    • 한국동물위생학회지
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    • 제43권1호
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    • pp.23-29
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    • 2020
  • Hepatitis E Virus (HEV) infection is a worldwide disease and the primary cause of acute viral hepatitis in the world. It can be isolated from many different species including pigs. HEV is a zoonotic pathogen and foodborne disease. The main animal reservoir is domestic pigs. It is usually asymptomatic in pig but it is a public health concern, causing acute hepatitis in humans of varying severity. This study focused on the presence of HEV in pig and pork product. One hundred feces and one hundred fifty serum samples were randomly collected from pigs in slaughterhouses in Gwangju from November in 2018 to February in 2020. In addtion, seventy-five pork products were collected from markets in Gwangju. Feces and pork product samples were examined for the presence of HEV RNA using an reverse-transcription realtime PCR (RT-qPCR) assay. Serum samples were tested for the presence of HEV-specific IgG antibodies using Enzyme-linked immunosorbent assay (ELISA). HEV antigen and antibody positive rates were 3.0% (3/100) and 19.3% (29/150), respectively, in Gwangju and nearby areas such as Jeonnam and Jeonbuk. However, HEV antigen was not detected from any of pork product in this study. In conclusion, the prevalence of HEV should be continuously monitored because HEV was sporadically detected in Gwangju and nearby areas.

교배형 분자마커를 이용한 신품종 밀리타리스 동충하초 '도원홍초 2호'의 품종 특성 (Varietal characteristics of new Cordyceps militaris 'Dowonhongcho 2ho' improved by mating type molecular markers)

  • 이병주;이미애;김용균;이순계;최영상;이병의
    • 한국버섯학회지
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    • 제15권3호
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    • pp.111-117
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    • 2017
  • 본 연구는 밀리타리스 동충하초의 교배형 유전자인 MAT1-1과 MAT1-2 두 종류에 특이적인 프라이머를 사용하여 multiplex PCR을 실시하였고, 그 결과 형성된 MAT1-1과 MAT1-2에 대한 233-bp와 191-bp에서 DNA 밴드를 통해 교배형 및 교배여부를 확인하였고 자실체 특성검정을 통해 품종특성이 우수한 새로운 동충하초 '도원홍초 2호'를 개발하였다. 신품종 '도원홍초 2호'의 자실체는 곤봉형이고 밝은 주황색을 띠었으며, 코디세핀 함량은 0.33%였고 자좌의 굵기와 길이는 각각 3.5 mm와 7.1 cm였다. '도원홍초'와 비교할 때, 새로운 '도원홍초 2호'의 수량은 7%가 증수되었고 경도가 높은 특징을 보였다. 균사생장의 적온은 $20{\sim}25^{\circ}C$였고 버섯 발생의 적온은 $18{\sim}22^{\circ}C$였으며, 접종에서부터 자실체 발생까지의 기간은 49.7일이 소요되었다. 신품종 '도원홍초2호'는 교배형 분자마커를 육종과정에 이용하여 효율성을 높였으며, 우수한 재배적 특성으로 동충하초 인공재배 및 산업적 생산에 기여할 것으로 판단된다.

배양 분리법을 통한 젓갈 내 원핵 세균 군집 분석 및 신규 미생물의 분리 (Analysis of Prokaryote Communities in Korean Traditional Fermented Food, Jeotgal, Using Culture-Dependent Method and Isolation of a Novel Strain)

  • 김민수;박은진;정미자;노성운;배진우
    • 미생물학회지
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    • 제45권1호
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    • pp.26-31
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    • 2009
  • 우리나라의 전통 발효 식품인 젓갈로부터 배양 분리법과 분자생물학적 분석법을 이용하여 원핵 세균 군집을 분석하고, 신규 미생물 분리를 목표로 하였다. 젓갈은 생산 지역과 주재료를 고려하여 17 종을 선정하였으며, 이들 젓갈 시료를 적정 희석배수로 희석하여 12종류의 미생물 선택배지에 도말, 배양한 후 나타난 집락(colony)을 형태학적 특성에 따라 무작위로 308개를 선정하여 분리하였다. 순수 분리된 미생물은 PCR 방법을 이용하여 16S rRNA 유전자의 염기서열을 분석한 후, 기존에 보고된 미생물 database와 비교함으로서 17종의 젓갈 내 미생물 군집을 확인하였다. 젓갈의 발효 및 숙성 과정에 관여하는 lactic acid bacteria (Leuconostoc 속, Weisella 속, Lactococcus 속, Lactobacillus 속, Carnobacterium 속, Marinilactibacillus 속, Tetragenococcus 속)와 Bacillus 속, Pseudomonas 속, Micrococcus 속, Brevibacterium 속, Microbacterium 속과 Kocuria 속이 17가지 젓갈에서 광범위하게 분리되었으며, Salinicoccus 속, Halomonas 속, Cobetia 속, Lentibacillus 속, Paracoccus 속, Psychrobacter 속이 소수 분리되었다. 또한 분리된 미생물의 계통학적 분석을 통하여 기존에 보고된 적이 없는 신규 미생물 14종을 분리하였다.

넙치(Paralichthys olivaceus) HSP90$\beta$ 유전자의 분자생물학적 연구 (Molecular Biological Studies on the Stress Protein HSP90$\beta$ Gene from Flounder (Paralichthys olivaceus))

  • 이재형;김영태
    • 한국미생물·생명공학회지
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    • 제32권4호
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    • pp.297-306
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    • 2004
  • 열 충격단백질(Heat shock protein : HSP)은 온도 스트레스에 대하여 세포 내에서 발현되는 단백질이다. HSP의 중요 분류군의 하나가 HSP90 family 이다. 여러 종류의 포유동물과 조류에서 HSP 유전자 특성에 대한 연구가 많이 진행되었다. 본 연구에서는 넙치(Paralichthys olivaceus)로부터 제조한 넙치 뇌 cDNA 유전자 은행을 이용하여 넙치 HSP90 cDNA 유전자를 분리하여 구성 염기서열의 특성을 밝혀 내었다. 염기서열의 분석결과 넙치의 hsp90$\beta$ 유전자는 2,791 개의 뉴클레오타이드로 구성되어 있고, 726개의 아미노산 잔기가 암호화되어 있었다. 넙치 hsp90$\beta$ 유전자는 European sea bass와 96.6% zebrafish와 92.9%, Atlantic salmon와 92.0%, 그리고 사람과는 89.5%의 염기서열 상동성을 지니고 있었다. 또한 HSP90 아미노산 서열을 바탕으로 척추동물 종들과의 진화계통수를 구축하였다. 넙치 hsp90$\beta$ 유전자의 mRNA의 분포 정도를 RT-PCR를 이용하여 조사하였다. hsp90$\beta$ 유전자는 조사한 모든 조직(뇌, 간, 신장, 근육, 비장)에서 높은 수준으로 발현이 되고 있었다. 또한, 넙치 hsp90$\beta$ 단백질을 대량발현하기 위하여 대장균에서 발현을 유도하였다.

혈액에서 Sucrose 양성 Vibrio vulnificus 분리 1예 (A Case of Sucrose-Positive Vibrio vulnificus Isolation from Blood Culture)

  • 김신무;송계민;김승아;최수연;임효빈;성치남
    • 대한임상검사과학회지
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    • 제36권2호
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    • pp.69-75
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    • 2004
  • Vibrio vulnificus is a halophilic bacterium frequently involved in human infection of seafood-associated primary septicemia and primary wound infection, mostly in men with over 40-years of age with underlying liver disease. The primary septicemia, which is the most common form of V. vulnificus infection in Korea, is defined as a systemic illness presenting fever or hypotension with recovery of V. vulnificus from blood or tissue without the apparent primary focus of infection. V. vulnificus typically do not produce acid from sucrose, but a case of primary septisemia was found in a patient at Chonnam K hospital in 1993 from whose blood a sucrose-fermenting strain was isolated. The patient was a 62-year-old man, heavy drinker, with underlying liver disease. He consumed a raw seafood dish two days before onset of the present illness. His symptoms were tenderness and swelling on the right foot. He rapidly developed septicemia, resulting in sudden death. V. vulnificus was isolated from the venous blood culture of the patient. On subculture, the isolate formed yellow colonies on TCBS and produced acid from sucrose. Because of these characteristics, species identification was not achieved by the API 20E and was delayed. Other characteristics of the isolate were identical to those of typical V. vulnificus. The isolate was common serotype O4A and possession of V. vulnificus-specific cytolysin gene was detected by PCR. The isolate was susceptible to all the antimicrobial agents tested including tetracycline, but was intermediate to colistin. In conclusion, it is important that microbiologists be aware of the presence of sucrose-positive V. vulnificus when he or she identifies gram-negative bacilli, which is isolated from the blood of patients with a recent history of raw seafood dish consumption.

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꽃노랑총채벌레 산란에 의한 포도 과피 달무리 반점: 종 특이적 분자진단법을 이용한 종동정과 반점 증상의 형태적 특징 (Halo Spot Symptom Induced by Oviposition of Frankliniella occidentalis on Grape Fruits: Molecular Diagnosis by a Species-specific DNA Amplification and Microscopic Characterization of the Symptom)

  • 안승준;조명래;박철홍;강택준;김형환;김동환;양창열
    • 한국응용곤충학회지
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    • 제53권3호
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    • pp.281-286
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    • 2014
  • 최근 국내 포도원에서 과실 표면에 구름모양의 흰색 얼룩이 자주 발견되고 있으나, 그 원인에 대해 명확히 알려진 바가 없어 방제대책을 세우지 못하고 있는 실정이다. 흰색의 달무리 반점은 포도 과실 표면에 부정형으로 퍼져 있으며, 그 중앙에는 총채벌레가 산란할 때 만들어진 작은 구멍의 상처가 남아 있다. 이 상처부위는 시간이 지나면서 코르크화되고 표피세포와 분리되어 딱지로 남거나 떨어져 나가게 된다. 이러한 증상은 총채벌레의 섭식이나 노린재의 흡즙에 의한 상처와는 구별된다. 산란구멍에서 발견한 총채벌레 알껍질에서 DNA를 추출하여 ITS2 부위의 염기 서열을 PCR-RFLP 방법으로 분자동정을 실시한 결과 꽃노랑총채벌레의 것으로 확인되었다. 미토콘드리아 COI 염기서열은 이러한 분자 동정 결과를 재확인하여 주었다. 본 연구결과는 포도 과피에 꽃노랑총채벌레 산란에 의해 유발되는 독특한 피해 증상에 대한 정확한 정보를 제공하며 포도원에서 이 해충에 대한 방제전략 수립에 도움이 될 것으로 여겨진다.

Isolation of an Rx homolog from C. annuum and the evolution of Rx genes in the Solanaceae family

  • Shi, Jinxia;Yeom, Seon-In;Kang, Won-Hee;Park, Min-Kyu;Choi, Do-Il;Kwon, Jin-Kyung;Han, Jung-Heon;Lee, Heung-Ryul;Kim, Byung-Dong;Kang, Byoung-Cheorl
    • Plant Biotechnology Reports
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    • 제5권4호
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    • pp.331-344
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    • 2011
  • The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.