• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.151-159
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• 2002

Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n＝9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.330-342
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• 2018

The Streptavidin and Biotin system has been studied most extensively as the high affinity non-covalent binding of Biotin to STR ($K_D=10^{-14}M$) and four Biotin binding sites in tetrameric Streptavidin makes this system useful for the production of multivalent antibody. For the application of this system, we cloned Streptavidin amplified from Streptomyces avidinii chromosome by PCR and fused to gene of hAY4 single-chain Fv antibody specific to death receptor 4 (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand. The hAY4 single-chain Fv antibody fused to Streptavidin expressed in Escherichia coli showed 43 kDa monomer in heated SDS-PAGE. However, this fusion protein shown in both non-heated SDS-PAGE and Size-exclusion chromatography exhibited 172 kDa as a tetramer suggesting that natural tetramerization of Streptavidin by non-covalent association induced hAY4 single-chain Fv tetramerization. This fusion protein retained a Biotin binding activity similar to natural Streptavidin as shown in Ouchterlony assay and ELISA. Death receptor 4 antigen binding activity of purified hAY4 single-chain Fv fused to Streptavidin was also confirmed by ELISA and Westernblot. In addition, surface plasmon resonance analysis showed 60-fold higher antigen binding affinity of the hAY4-STR than monomeric hAY4 ScFv due to tetramerization. In summary, hAY4 single-chain Fv fused to Streptavidin fusion protein was successfully expressed and purified as a soluble tetramer in E. coli and showed both Biotin and DR4 antigen binding activity suggesting possible production of bifunctional and tetrameric ScFv antibody.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.67 no.1
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• pp.9-16
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• 2022

Rice is one of the most important staple foods in Wnju, Jeonbuk, South Korea. However, rice consumption has dramatically decreased as eating habits have diversified owing to rapid economic growth. Recently, floury endosperm rice varieties have been developed to invigorate the rice processing industry, because dry-milled rice flour is economically and environmentally suitable for massive rice flour distribution. The National Institute of Crop Science has developed 'AromaT', an early-maturing black rice with floury-endosperm, suitable for tea and dry milling. 'AromaT' was derived from a cross between 'Suweon542' as the floury endosperm source and 'Heugjinju' as the black and aromatic source. In this study, 'AromaT' and its parents, 'Suweon542' and 'Heugjinju', were analyzed for agronomic traits, anthocyanin content, and their major physicochemical properties by different planting date. The field experiment was conducted in Wanju, Jeollabuk-do Province, South Korea, in 2019. The transplanting dates were May 30 (ordinary season), June 25 (double-cropping season), and July 10 (late season). The yield performance of brown rice 'AromaT' was 330 kg/10 a in the double-cropping cultivation method and was the highest among the transplanting dates. The floury endosperm of 'AromaT' was derived from 'Suweon542' containing 'flo7', located on chromosome 5 and known to control floury endosperm. With the late planting date, the anthocyanin content of 'AromaT' was 570.5 mg/100 g, much higher than that of 'Heugjinju' (376.3 mg/100 mg). The brown rice of 'AromaT' also exhibited the pop-corn-flavoring 2-acetyl-1-pyrroline, exclusively detected in aroma rice varieties. The average particle sizes of 'AromaT' and 'Suweon542' were 67.12 ㎛ and 70.9 ㎛, respectively, lower than that of 'Heugjinju' (95.5 ㎛) with a black transparent endosperm. The average damaged starch content of 'AromaT' was 8.1%, lower than that of 'Heugjinju' (10.05%) and Suweon542 (9.5%). As a result, 'AromaT' with high anthocyanin content, fine particle size, and low damaged starch content is expected to provide a new rice material in various processing fields.