• Title/Summary/Keyword: reverse mapping

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Analysis of Antigenic and Genetic Variability of G-protein of Respiratory Syncytial Virus Subgroup A Isolated in Korea over 8 Years(1990~1998) (국내에서 분리된 Respiratory Syncytial Virus A 아군의 항원성의 변이와 G-단백 mRNA의 RT-PCR 생산물의 제한효소 처리 및 염기 서열 결정을 통한 유전자 변이의 분석)

  • Choi, Eun Hwa;Park, Ki Ho;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.219-233
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    • 1999
  • Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

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Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.