• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.17-25
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• 2013

The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

• Pediatric Infection and Vaccine
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• v.25 no.2
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• pp.72-81
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• 2018

Purpose: This prospective study aimed to investigate the therapeutic efficacy of lamivudine in children with chronic hepatitis B virus (HBV) infection. Methods: During July 2003 through October 2015, children with chronic hepatitis B who visited our institution were included in this study. Fifty-five patients, who received first-line treatment of lamivudine (3 mg/kg, 100 mg maximum) for over three months, were enrolled. After initiating lamivudine, alanine aminotransferase (ALT), HBV-DNA, and HBV markers were followed up at 1 month, 3 months, and every 3 months, thereafter. The treatment endpoint was determined as 1) normalization of ALT, 2) HBeAg seroconversion, and 3) anti-HBe positivity for twelve consecutive months. Results: Thirty-one male (56.4%) and 24 female (43.6%) patients were included. The mean age at treatment initiation was 8.1 years. The mean duration of treatment was 23.4 months. ALT normalization was found in 98.2% (54 of 55). Anti-HBe seroconversion was found in 70.6% (36/51). Loss of HBsAg was found in 10.9% (6/55). All biochemical responses occurred under age seven. The rate of virologic response (defined as HBV-DNA <2,000 IU/mL) at six months after treatment initiation was 78.7% (37/47). At twelve months after reaching treatment endpoint, 87.2% (34/39) maintained their virologic response. Resistance to lamivudine was found in 16.4% (9/55). Conclusions: Lamivudine treatment in Korean pediatric patients with chronic hepatitis B showed better outcomes compared with other studies that implemented similar protocols in foreign populations. Further studies are needed to investigate the efficacy of newly recommended antiviral drugs on the Korean pediatric population.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.225-234
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• 2006

In the present study toxicity and immunostimulating activity of the lectin(KML-C), which was extracted from Korean mistletoe(Viscum album coloratum) were investigated in swine. To determine the toxicity, lectin was injected into thigh or cervical muscles of 4-week-old piglets(Landrace) and observed clinically and pathologically. For determination of the immnunostimulating activity, lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine of Aujeszky's disease virus(ADV)(NYJ1-87) which was inactivated by 0.2% formalin was injected into the cervical muscle of antibody-negative piglets in the same age group. Subpopulation of the immune cells and serum neutralizing(SN) antibodies in the piglets were examined after vaccination, and resistance of the piglets against challenge by virulent NYJ1-87 was further examined. The results were also compared with those from piglets injected with aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccine of inactivated NYJ1-87 and NYJ1-87 vaccine without adjuvant, and the results are as follows. By injection of lectin with $30{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets died after signs such as dyspnea, fever, systemic erythema and subcutaneous hemorrhages, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with $7{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets showed signs such as edema and cutaneous hemorrhage in the injected area, lameness and depression, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with 1, 3 and $5{\mu}g/kg$ of body weight to the thigh muscle of each one piglet, signs such as congestion, induration and grayish coloration in the injected area, depression and inappetence were observed in all piglets. Toxic changes were also observed in the liver and kidney of piglets by lectin of 3 and $5{\mu}g$. By injection of lectin with 0.5 and $0.7{\mu}g/kg$ of body weight to the cervical muscle of each 9 piglets, all piglets were clinically normal and there were no significant changes in blood counts and chemistry values. Whereas, epithelial swelling and vacuolation of convoluted tubules were observed from one piglet injected with lectin of $0.7{\mu}g$, and necrosis and fibrosis of muscular fiber were observed in the muscle of one piglet injected with lectin of $0.5{\mu}g$. Only population of sIgM+ B lymphocytes increased among immune cells in all of 15 piglets immunized with lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine, while compared to those in $Al(OH)_3$-adjuvanted vaccine and vaccine without adjuvant. No additional stimulation to the immune cells was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. In piglets immunized with lectin-adjuvanted vaccine, SN titers in reciprocal values for loge were 1.3-4.0 at 1-4 weeks after vaccination, which was similar to those with 1.0-3.3 by vaccine without adjuvant but lower than those with 2.0-5.7 by $Al(OH)_3$-adjuvanted vaccine. Also, no additional increase in the SN titers was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. Piglets immunized with lectin-adjuvanted vaccine were resistant to challenge by the virulent NYJ1-87 at 4 weeks after vaccination, and the SN titers reached to 5.0 one week after challenge, which was higher than those with 4.0 by vaccine without adjuvant but somewhat lower than those with 7.7 by $Al(OH)_3$-adjuvanted vaccine.

• Korean journal of applied entomology
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• v.25 no.2 s.67
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• pp.113-120
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• 1986

Soybeans collected from different locations in Korea were planted for tests of seed transmission of soybean mosaic virus (SMV). The percentage of seed transmission ranged from 20.5 to 29.5% in 14 seedlots including soybean cultivar Namhe and 12 to 19% in other 44 seedlots. However, no seed transmission of SMV was observed in soybean cultivar Younkiyongho. SMV was detected from embryo and cotyledon of soybean seeds. The infection of SMV was highly detected from premature seeds than from fully mature seeds, and higher from seeds harvested from plants infected before June 20 than plants infected around July 20 and August 20. No significant relationship was observed between seed transmission of SMV and mottling of seeds. The incidence of soybean mosaic disease followed by one month after peak of aphid population. The number of aphids was less on leaves of soybeans with short and dense trichomes whereas it was higher on leaves of soybean with long and sparse trichomes. Generally, the number of aphids was decreased on leaves with long and dense trichomes as the growth progressed. Soybean cultivar Columbus and 14 cultivars were susceptible, Chief and 14 cultivars were moderate, but Jangbaek and 17 other cultivars were resistant to SMV when inoculated with one isolate of SMV.

• 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 Breeding Science
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• v.40 no.3
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• pp.328-331
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• 2008

Hwangeumnodeul is a new japonica rice cultivar developed from a cross between Milyang165 and HR15151-B-21-3 at Honam Agricultural Research Institute (HARI), NICS, RDA, in 2007. This cultivar has short grains and about 139 days of growth duration from direct seeding to harvesting in the southern plain areas. Culm length of this cultivar is 78 cm and the number of spikelet per panicle is similar to that of the check cultivar, Nampyeongbyeo, while filled grain rate is lower than the check variety. This cultivar is moderately resistant to leaf blast, bacterial blight pathogens of $K_1$, $K_2$ and $K_3$ and stripe virus but susceptible to major virus diseases and insect pests. Milled kernel of Hwanggeumnodeul is translucent with non-glutinous endosperm. This cultivar has about 5.7% and 18.6% of protein and amylose contents, respectively. Palatability of cooked rice of Hwanggeumnodeul is similar to Nampyeongbyeo. Its milling recovery and head rice were 75.6% and 90.0%, respectively. Hwanggeumnodeul yielded 5.69 and 5.61 MT/ha in milled rice at the under wet-direct and dry-direct seeding conditions, respectively which are 17% and 12% higher than the check variety, Juanbyeo. It yielded 5.86 MT/ha, 8% higher than Nampyeongbyeo at the standard fertilizer level of the ordinary transplanting cultivation. "Hwanggeumnodeul" would be adaptable for ordinary transplanting as well as direct seeding in the southern plain including Chungcheong province.

• Korean Journal of Breeding Science
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• v.43 no.3
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• pp.201-205
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• 2011

'Hangangchal 1' is a new glutinous rice cultivar of second generation Tongil-type with a mid-maturing ecotype that developed by the rice breeding team of Yeongnam Agricultural Research Institute (YARI), RDA. in 2006. This cultivar was derived from a cross between 'Hangangchal', a Tongil-type glutinous cultivar and 'YR8208-20', a high yield potential in 1986/1987 winter season. 'Hangangchal 1' was selected by pedigree and bulk breeding methods from $F_3$ to $F_6$ populations. A promising line, YR10498-8-1-3, was selected and designated as 'Milyang 167' in 1997. The local adaptability test of 'Milyang 167' was carried out at seven locations during 3 years in 1998, 2005, and 2006. It has tolerance to lodging with good canopy architecture as 87cm of culm length. This cultivar is resistant to bacterial blight $K_1$, $K_2$, and $K_3$ race, rice stripe virus, rice dwarf virus, and leaf blast disease. The milled rice endosperm of 'Hangangchal 1' is glutinous and its whiteness was almost similar compared to 'Shinseonchalbyeo'. The yield of milled rice of 'Hangangchal 1' was average 5.97 MT/ha at ordinary cultivation of 9 kg/10a N fertilizer level in local adaptability test. This cultivar would be adaptable to the mid and southern plain of Korea.

• Korean Journal of Breeding Science
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• v.41 no.2
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• pp.163-167
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• 2009

Jogwang is a new early maturing japonica rice developed in 2007 from a cross between Milyang187 and YR21113-B-B at the Department of Functional Crop Science, NICS, RDA. This cultivar is very suitable to the rice-cash crop double cropping system. Heading date of Jogwang is 2 days earlier than Keumobyeo under the late transplanting cultivation on July 10 at the Yeongnam plain. The tolerance level of this variety to leaf discoloration at seedling stage is very similar to Keumobyeo. It showed slightly lower viviparous germination and premature heading. This cultivar showed resistant reactions to leaf blast and rice stripe virus disease but susceptible to bacterial blight disease and major insect pests. The ratio of milling and head rice recovery of Jogwang is 76.5% and 64.5%, respectively. The milled kernels are translucent with non glutinous endosperm. This cultivar has 7.3% protein and 18.5% amylose content. In local adaptability test, showed that the milled rice yield of Jogwang is $4.90\;MT\;ha^{-1}$. This cultivar is suitable for planting in the plain paddy fields of Honam and Yeonnam regions in Korea.

• Journal of Plant Biotechnology
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• v.34 no.1
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• pp.11-17
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• 2007

Previously developed transgenic watermelon rootstocks (gongdae) inserted by CGMMV-CP were examined to test the virus tolerance levels. In the restricted plastic house, the $T_{3}$ watermelon rootstock showed tolerance to CGMMV until 70 days after inoculation on the leaves while the non-transformed watermelon rootstock became susceptible at 20 days after inoculation. In the field, tolerance efficiency of transgenic rootstocks maintained up to 40% at 71 days after contamination with CGMMV in the soil while all of the non-transformed rootstocks became susceptible at 37 days with the same condition. In the same field, transgenic rootstocks showed more tolerance to CGMMV than the non-transformed rootstocks as those were inoculated on the leaves, but it showed only 10 days delay before being susceptible. Therefore, transgenic rootstocks have a characteristic of delay effect against CGMMV susceptibility, rather than resistance character. From $T_{3}$ rootstocks homozygous for the CGMMV-CP horticulturally favorable individuals were selected for further breeding and a transgenic line was finally obtained at the $BC_{1}T_{5}$. A material transfer experiment was conducted to find out if the DNA, RNA or expressed protein in the transgenic rootstocks could move to the grafted scion (non-transformed watermelon, Super-Kumcheon). PCR, northern, and western blot analysis were performed and no evidence of transferring of those materials from rootstock to scion was ever found.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.175-182
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• 1995

Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.