• Title/Summary/Keyword: Coat Protein

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Expression of a Functional Anti-Cucumber Mosaic Virus Single-Chain Variable Fragment Antibody in Tobacco Plants (Nacotiana tabacum)

  • Heng Chua Kek;Khalid Norzulaani;Othman Retina Yasmin
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.9-14
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    • 2006
  • As an alternative method to produce low cost reagents for immunodiagnosis and protect the plants from viral disease, a gene encoding a single chain variable fragment(scFv) recombinant antibody targeted to the coat protein of cucumber mosaic virus (CMV) was expressed in Nacotiana tabacum. The source of the scFv recombinant antibody gene was from spleen tissue of an immunized mouse. The gene was initially cloned into the pCANTAB5E phagemid and expressed in E. coli. In the following study, the antibody gene was subcloned into the plant expression vector, pCAMBIA-1301 and introduced into tobacco leaf tissue via Agrobacterium tumefacients mediated transformation. After transformation, 56 out of 58 plants were shown to carry the desired anti-CMV scFv gene by PCR analysis. Overall, only 12.5% of the 56 putative transgenic plants were found to express the antibody to a detectable level.

Detection of Lily Symptomless Virus Using RT-PCR Technique (RT-PCR 기법을 이용한 Lily Symptomless Virus의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.187-190
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    • 1996
  • 백합으로부터 total RNA를 분리하여 LSV 외피단백질 유전자의 551 bp에 해당하는 특정 염기서열을 증폭할 수 있는 primer로 RT-PCR를 수행하였다. 그 결과 Lilium oriental hybrid cvs. Miani, Marco Polo, Casablanca, Le Reve 품종에서 551 bp의 DNA 절편이 증폭되었고 이 절편의 염기서열을 분석한 결과 LSV외피단백질 유전자의 일부임을 확인할 수 있었다. 그러므로 RT-PCR 방법으로, 실험에 사용하 s4품종 모두 LSV에 감염되어 있음을 알 수 있었다.

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Determination of Nucleotide Sequences of cDNA from Cucumber Mosaic Virus-As RNA4 (As계의 오이 모자이크 바이러스 RNA4의 염기서열 결정)

  • 김상현;박원목;이세영;박영인
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.176-181
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    • 1996
  • Aster yomena로부터 분리한 오이 모자이크 바이러스(cucumber mosaic virus) (CMV-As)의 RNA4로부터 완전한 길이의 cDNA를 합성하고 그 전체적인 염기서열(1,043 nt`s)을 결정하였다. CMV-As RNA4는 73개의 염기로 구성된 5`말단의 leader 부위, 657개의 염기로 구성된 외피단백질(coat protein) 유전자 부위 및 312개의 염기로 구성된 3` 말단의 비번역 부위로 구성되어 있음을 확인하였다. 외피단백질 유전자 부위의 염기서열을 다른 계통의 CMV와 비교해 볼 때 그 염기서열이 보전적으로 존재하고 있으나 그 외의 부분은 다양함을 확인하였다. 특히 3` 말단부위의 61개의 염기로 구성된 부위(959-1019)는 다른 계통의 CMV에서는 상당히 유사하지만 CMV-As도 다른 CMV처럼 tRNA와 유사한 구조를 역시 형성함을 확인하였다. CMV-As의 RNA4 염기서열을 다른 계통의 CMV와 비교할 때 CMV-I17F와 가장 유사하였으며(91.9%) S형의 CMV-M과는 가장 낮은 동일성을 보였다(71.1%). 외와 같은 염기성열의 비교 결과와 EcoRI 제한효소 인식부위의 존재로 미루어 CMV-As는 WT형으로 분류된다.

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Story of Johnsongrass Mosaic (Poty)virus in Australia

  • Oh, Hae-Young
    • Journal of Life Science
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    • v.12 no.2
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    • pp.61-74
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    • 2002
  • One of the major aims in studying plant viruses is to minimise the development of symptoms in infected plants. With the advent of in vitro transcript mediated research on plant viruses, substantial progress has been made. This article describes the biology of a plant specific RNA virus, Johnsongrass mosaic virus (JGMV), important to Australian sorghum and corn agriculture and, in particular, at a molecular level which of the RNA sequences in its genome that make it possible for the virus to move from cell to cell, and eventually spread systemically throughout the entire plant. The JGMV has caused considerable yield losses in maize and sorghum over a number of years in Australia. Incidents where 100% of the crop has been infected are on record. The use of this virus is convenient under laboratory conditions because it can be readily transmitted by mechanical inoculation with infected leaf sap, which obviates the need for maintaining aphid colonies. The JGMV is a single stranded positive sense RNA virus.

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Identification and Characterization of Tobamoviruses Isolated from Commercial Pepper Seeds (시판 고추 종자에서 분리한 Tobamovirus의 동정 및 특성 조사)

  • 한정헌;손성한;나용준
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.164-169
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    • 2001
  • Two Tobamoviruses showing different local lesion types on Nicotiana glutinosa was isolated from commercial pepper seeds. These viruses were designated Tobamovirus-6 (T-6) and Tobamovirus-19 (T-19). The biological and serological assays revealed that T-6 and T-19 were closely related to Pepper mild mottle virus (PMMoV) and Tomato mosaic virus (ToMV), respectively, The isolates also had low similarity in the array of viral coat protein gene sequences, of which T-19 was most identical to known strains of ToMV, while T-6 was closely related to PMMoV.

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Diagnosis of Potato Leafroll Virus with Digoxigenin-labeled cRNA Probes (Digoxigenin으로 표지된 cRNA 프로브를 이용한 감자잎말림바이러스(PLRV)의 짐단)

  • 서효원;함영일;오승은;신관용;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.636-641
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    • 1998
  • Digoxigenin (DIG) was used to prepare nucleic acid probe for the detection of RNA of potato leafroll virus (PLRV) in the potato leaf extracts. The 0.6 kb coat protein (CP) gene cDNA of PLRV in plasmid pSPT 18 vector was labeled with digoxigenin by in vitro run-off transcription and then used for cRNA probe. In the several buffers tested for increase the total RNA extraction efficiency AMES buffer was the most suitable for this detection method. The RNA extracts from potato leaves shown symptoms of PLRV were dot blotted onto nylon membrane and hybridized with labeled RNA probes. After hybridization, labeled RNA bound to PLRV RNA on membrane was detected with anti-digoxigenin alkaline phosphatase. 5-bromo-4-chloro-3-indolyl-phosphate/nitroblue tetrazolium (NBT) salt and CSPD were used as substrate for colorimetric and film exposure detection, respectively. These detection methods were very sensitive allowing for detection of 1/32 diluted total RNA extract from 100 mg leaf tissue.

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Effects of Plant Growth Regulators on Physiology of Germinating Punax ginseng Seed (식물 생장조절제가 인삼 (Punax ginseng) 종자의 발아생리에 미치는 영향)

  • Eung-Ryong Son;Won-Mok Park;C. Pertzsch
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.24 no.1
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    • pp.99-106
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    • 1979
  • The undehiscent ginseng seed did not germinate, even if the seeds were treated with GA_3, kinetin or IAA. Only GA_3 stimulated germination of dehiscent ginseng seed. The physiological roles of gibberellic acid on stimulation of the seed germination were enhancing production of soluble carbohydrate and sucrose. Then gibberellic acid stimulated biosynthesis of insoluble cellural materials and amino acids from sugars and incorporation of amino acids into protein. The fruit coat of ginseng seed did not impede water imbibition, but did function as water absorbor and reservoir.

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Detection of Cucumber Mosaic Virus by RT-PCR Using a Simple and Rapid Crude Sap Extraction Method (간이 조즙액 추출법을 이용한 RT-PCR 방법에 의한 오이 모자이크 바이러스의 검정)

  • 이상용;홍진성;이진상;최장경
    • Korean Journal Plant Pathology
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    • v.12 no.4
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    • pp.432-436
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    • 1996
  • 역전사 중합효소 연쇄반응(RT-PCR)을 이용하여 담배(Nicotiana glutinosa)에 증식시킨 7종의 오이 모자이크 바이러스(CMV)를 검정하였다. RT-PCR을 위한 간단하고 신속한 바이러스 핵산의 조즙액 추출법이 개발되었으며, CMV 외피단백질 유전자 부위를 기초로 하여 제작한 20개의 염기로 구성된 primer를 사용하여 RT-PCR을 실시한 결과, 약 490 염기쌍의 DNA 단편들이 이병식물의 조즙액으로부터 증폭되었다. EcoRI 및 MspI을 이용한 RT-PCR 산물의 분석에 의하여, 공시한 7종의 바이러스는 모두 CMV subgroup I으로 동정되었다. Ouchterlony 한천젤 이중 확산법을 이용한 항혈청 검정에서도 7종의 바이러스 모두 CMV-Y의 항혈청과 단일의 침강선을 형성하였다.

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Characterization of Carnation mottle camovirus(CarMV) Isolated from Lilium spp. in Korea

  • Park, J.H.;J.H. Sung;H.Y. Shin;M.U. Chang;S.N. Yoo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.150.2-150
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    • 2003
  • Carnation mottle carmovirus(CarMV) was isolated from Lilium spp. in Korea. This isolate, CarMV, was done bioassay, which plants were Dianthus caryophyllus, Gomphrena globosa, Chenopodium amaranticolor, Dianths chinensis. CarMV was propagated on the leaves of Chenopodium amaranticolor with the crude-sap inoculation method and purified by Mossops method(1976). We produced antiserum against CarMV and analyzed the antiserum specificity with ELISA, Gel diffusion method, and Rapid Immunofilter Paper Assay (RIPA). From these results of the assay, RIPA method was simple and rapid for CarMV detection. We have established successfully the CarMV detection system. CarMV coat protein gene was amplified by RT-PCR with specific primers and sequencing analysis was done.

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Variation of Protein Content and Amino Acid Composition in Perilla Germplasm (들깨 유전자원의 단백질함량과 아미노산조성)

  • Lee, Jung-Il;Bang, Jin-Ki;Lee, Bong-Ho;Kim, Kwnag-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.5
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    • pp.449-463
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    • 1990
  • To obtain the basic informations on quality improvement, seed protein and amino acid composition were analyzed in 460 strains of perilla germplasm. Among the tested strains, total protein content ranged from 17.9% to 28.1 % with the 23.6% of varietal means. Form the experiment, Namji, Sandong, and Eunjin were selected as high protein strains of which content was as high as 28.1%. In protein content, collected strains from Jeonnam province showed highest, and was not significantly different by maturity, but this characteristics showed differences by seed coat color and 1,000 seed weight. The significantly negative correlation was observed between protein content and seed setting ratio. However it was observed that significant and high positive correlation between protein and oil content. A calibration for an Infra-Alyzer 450 using log reflectance readings at 2208, 1982, 1940 and 1722nm could be used without adjustment for the measurment of the protein content in perilla with a standard deviation of differences against micro-kjeldahl of 0.27%. The amino acid composition of perilla was similar to the other oilseed crops, and showed a relatively high lysine and methionine content. Further, amino acid composition of perilla seed was exellently characterized with bal ance and higher than FAO recommendation. Major amino acids were indentified as a glutamic acid and arginine in perilla seed protein.

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