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Kwak, Bo Yeon,Shon, Dong Hwa,Kwon, Byung Joon,Kweon, Chang Hee,Lee, Ke Ho 한국미생물 · 생명공학회 2001 Journal of microbiology and biotechnology Vol.11 No.1
Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contarninated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellular polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1㎎/ml. Among the 59strains tested (including 18species of Aspergillus, 16 of Penicilliun, 11 of Fusariun, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and Penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1 GI 1 was on the carbohydrate moiety when 1 to 100 ㎍/g A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice or potato (r=0.91-0.99).
Detection of Fusarium Species by Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibody
KWAK, BO-YEON,KWON, BYUNG-JOON,KWEON, CHANG-HEE,SHON, DONG-HWA 한국미생물 · 생명공학회 2003 Journal of microbiology and biotechnology Vol.13 No.5
Enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of Fusurium species, known as harmful fungi in food. One of the hybridoma cell lines (1B8) which produced a monoclonal antibody (Mab) specific to Fusarium extracellular polysaccharide (EPS) was screened and the Mab was produced and purified. A detection limit of the sandwich ELISA against F: moniliforme EPS was 0.001 ㎍/ ml in the standard curve. Among the 59 strains tested, most Fusarium species showed high reactivity with Mab I B8, even when the culture broths were diluted 100,000 times. On the other hand, the other genera, except A. versicolor and Trichodennu viride, had no reactivity. When 1 to 100 pg/g of F: monilifonne EPS was spiked into rice, potato, and mandarine orange, the average recoveries were 151 %, 84%, and 94% respectively, determined by sandwich ELISA. The correlation coefficients between the EPS levels determined by sandwich ELISA and the dry mycelial weight of the liquid culture of F. moniliforme, as well as between the EPS and colony forming unit in solid culture of potato, were 0.97 and 0.91, respectively.
KWAK, BO-YEON,KWON, BYUNG-JOON,KWEON, CHANG-HEE,SHON, DONG-HWA 한국미생물 · 생명공학회 2004 Journal of microbiology and biotechnology Vol.14 No.2
The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELISA) system was developed in order to simultaneously detect the extracellular polysaccharide (EPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lGll alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.
Jong-Hee Lee,Yeon-Jae Hur,Do-Yeon Kwak,Jun-Hyun Cho,Yeong-Nam Yoon,Bong-Choon Lee,Ji-Yun Lee,Sang-Yeong Kim,Yeong-Bo Sohn,Un-Sang Yeo,Yu-Chun Song,Choon-Woo Lee,Min-Hee Nam,Jae-Keun Sohn 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
The green rice leafhopper (GRH), Nephotettix cincticeps Uhler, is one of the most serious insect pests affecting cultivated rice (Oryza sativa L.) in temperate regions of East Asia. To understand the genetic basis of the GRH resistance, a F2 population derived from across between a highly resistant variety,Cheongnam and a susceptible variety, Junambyeo was analyzed by genetic analysis and association mapping. GRH resistance was evaluated using the F2 populations. The results showed that a single dominant gene in Cheongnam. DNA from 22 F2 individuals being either resistant or susceptible were pooled to produce bulk resistant and bulk susceptible DNA samples. Parents and bulks were screened with 192 SSR markers and twolinked SSRmarker, RM6082 and RM20145 were identified.Subsequent mapping in the original mapping population showed that thelocusis flanked by the SSR markers, RM20130 and RM20152 on chromosome 6. To physically map this locus, the-linked markers were landed on the artificial chromosome clones of the reference cv., Nipponbare, released by the International Rice Genome Sequencing Project. The DNA markers found to be closely linked to Grh3 would be useful for marker-assisted selection for the improvement of resistance to GRH in rice.