• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.92-97
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• 2012

Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1-16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

• Journal of Mushroom
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• v.2 no.3
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• pp.115-126
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• 2004

Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.2
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• pp.79-88
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• 1998

This experiment was conducted from May to August in 1997 to selected the wrn hybrids being suitable for silage at farm in the Kongju National University through the comparison of growth characteristics, forage yield and growth analysis about native and imported corn hybrids for silage production. In this experiment, trial design was a randomized block design with three replication, testing varieties were 4 hybrids (Suwon 19, Kwanganok, Whengsungok, Suwonok ) of native corn hybrids and 13 hybrids (P 3156, P 3352, P 3144w, DK 501, DK 689, DK 713, DK 729, H 643.99, H 545.64, H 645.12, HC 7466, H 644.18, H ALISEO) of imported corn hybrids. The results obtained are summarized as follows; 1. The emergence rate of H643.99 was the highest with 97.0%. In rice black streaked dwarf virus(RBSOV), the hybrid of HC 7466 was lower infected with 1.6% than other hybrids. The plant hight of P 3144w was the highest with 339 cm and the stem length of P 3156 was the highest with 261 cm. In native com hybrids, the plant height and stem length of Kwanganok were recorded with 306 cm and 235 cm, respectively. 2. Leaf number and leaf area of Kwanganok were the greatest with 16 sheet per plant and $5,180\;{\textrm{m}^2}/l0a$, respectively. H 645.12 and H 545.64 had the greatest in ear to total dry matter ratio with 49.5% and 49.4%, respectively. 3. The fresh matter yield was significantly difference between growth stage, So Suwon 19 had the most level at 15 days before silking, P 3352 had the most level at silking date, Kwananok had the most level at 35 days a after silking. The fresh matter yield of native com hybrids such as Suwon 19 and Kwanganok was not apparent diffreences as compared with imported corn hybrids. 4. As the results of survey with dry weight, the quantity of dry matter accumulation were increase after silking. The varieties of P 3352, P 3156, Kwanganok, OK 713 were more quantity of dry matter production than DK 501, HC 7466. The Kwanganok of native com hybrid and Pioneer strain with high percentage of dry matter were higher dry weight than Limagrain strain. 5. HC 7466 had the largest LAR with $6.53\;{\textrm{cm}^2}/g$, H545.12 had the lowest LAR with $3.30\;{\textrm{cm}^2/g}$. P 3144 had the largest LAI, DeKalb strain including DK 713 were larger apparently than Limagrain strain including HC 7466 with 3.15. 6. The RGR of testing varieties was little difference of statistical significantly, but DK 501, and HC 7466 were lower than other corn hybrids. The CGR of native and American varieties was no apparent differences, but that of Limagrain strains were a large variation. According to the results obtained by this experiment, the eary growth such as emergence rate and RBSDV infection rate of Limagrain strains was more excellent than other strains. P 3156, P 3352, P 3144w, DK 713 and HC 7466 were suitable for silage condition such as dry matter yield, percentage of dry matter and % ear to total dry matter. The fresh and dry matter yield of native corn hybrids such as Suwon 19 and Kwanganok were not apparent differences as compared with imported corn hybrids, but percentage of dry matter was lower than other imported corn hybrids.

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.143-151
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• 1997

Interspecific hybrids between Aspergillus niger and Penicillium notatum (Tyr-), hyperlipolytic enzyme-producing fungi, were obtained by nuclear transfer technique. Optimal conditions for formation of intergeneric hybrids were investigated. Maximum production of protoplasts was obtained by 1% Novozyme 234 at $30^{\circ}C$ for 3 hrs and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6 M KCl. Frequencies of hybrid formation by nuclear transfer were $3.8{\times}10^{-3}{\sim}1.3{\times}10^{-3}$. From the observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyotypes are aneuploid. The hybrids showed $1.2{\sim}1.7$ fold higher lipase activities than parental strains. It was strongly supported by results of this study that nuclear transfer technique is much more efficient in the formation of intergeneric hybrids than protoplast fusion and is very useful for the improvement of strains.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.4 no.2
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• pp.155-160
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• 2001

Purpose: Helicobacter pylori has been known to have diverse vacA allelic types. The purpose of the study was to identify vacA diversity in Korea and design new primers for signal sequence alleles indigenous to Korea. Methods: Fifty antral biopsy specimens, which had been proven to be H. pylori-positive, were examined for vacA status; signal sequence and mid-region. After PCR amplification and DNA sequencing, vacA alleles of Korean H. pylori strains were compared with those from other countries. Results: Among Korean H. pylori strains vacA alleles with all combinations of signal sequence and mid-region were found, with the exception of s1b or s2. vacA genotype s1c/m1 was predominant in Korea. We found that GGGAGCGTTR in s1a and GGGGYTATTG in s1c were the indigenous sequences to Korea and constructed the new Korean specific primers for the vacA signal sequence; VASK-F, VASK-R, S1AK-F, and S1CK-F. Conclusion: This study showed that s1c/m1 is the predominant type of vacA allele in Korea. We designed new primers for the vacA signal sequence.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.13-13
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• 2014

Two kinds of mushrooms, Gumji (金芝; Ganoderma) and Soji, were described in old book of Samguksagi (History of the three kingdoms; B.C 57~A.D 668; written by Bu Sik Kim in 1145) in Korea-dynasty. Many kinds of mushrooms were also described in more than 17 kinds of old books during Chosun-dynasty (1392~1910) in Korea. Nowadays, mushroom cultivation has been increased through out the world last decade years. Production of mushrooms has also been increased 10-20% and many varieties have been cultivated. Similar trends were also observed in Korea. Approximately two hundred commercial strains of 37 species in mushrooms were developed and distributed to cultivators. Somatic hybrid variety of oyster mushroom 'Wonhyeong-neutari' were developed by protoplast fusion, and distributed to grower in 1989. The fruiting body yield index of somatic hybrids of Pleurotus ranged between 27 and 155 compared to parental values of 100 and 138. In addition, more diverse mushroom varieties such as Phellinus baumi, Auricularia spp., Pleurotus ferulae, Hericium erinaceus, Hypsizigus marmoreus, Grifola frondosa, Agrocybe aegerita and Pleurotus cornucopiae have been attempted to cultivate in small scale cultivation. Production of mushrooms as food was 190,111 metric tons valued at 800 billion Korean Won (one trillion won if include mushroom factory products; 1dollar = 1,040 Won) in 2011. Major cultivated species are Pleurotus ostreatus, Pleurotus eryngii, Flammulina velutipes, Lentinula edodes, Agaricus bisporus, and Ganoderma lucidum, which cover 90% of total production. Since mushroom export was initiated from 1960 to 1980, the export and import of mushrooms have been increased in Korea. Technology developed for liquid spawn production and automatic cultivation systems lead to the reduction of the production cost resulting in the increasement of mushroom export. However some species were imported because of high production cost for these mushrooms requiring the effective cultivation methods. Developing of effective post-harvest system will be also directly related to mushroom export. In academic area, RDA scientists have been conducting mushroom genome projects. One of the main results is the whole genome sequencing of Flammulina velutipes for molecular breeding. An electrophoretic karyotype of of F. velutipes was obtained using CHEF with 7 chromosomes, with a total genome size of approximately 26.7 Mb. The mususcript of the genome of F. velutipes was published in PLOS ONE this year. For medicinal mushrooms, we have been conducting the genome research on Cordyceps and its related species for developing functional foods using this mushroom. In 2013, Korea Food and Drug Administraion (KFDA) approved Cordyceps mushroom for its value as an immune booster.

• Journal of Mushroom
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• v.14 no.3
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• pp.81-89
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• 2016

White button mushroom (Agaricus bisporus) is the most widely consumed mushroom in the world, and domestic consumption is increasing owing to expansion of the westernized diet in recent years. Before 2000, most Korean mushroom farmers cultivated foreign varieties, but recently, Korean breeders are developing hybrid strains using molecular breeding tools. To produce a better mushroom cultivar, we evaluated some traits of button mushroom Agaricus bisporus strains. Mycelial growth rate at different culture temperatures was investigated by 25>20>15>30>$10^{\circ}C$. Recently developed domestic varieties had superior mycelial growth rate compared to the traditionally grown species. In particular, the ASI1338 strain showed excellent mycelial growth rate at different temperatures. Further, we confirmed that ASI1007, ASI1085, ASI1310, ASI1339 were strongly resistant to bacterial brown blotch, whereas ASI1053, ASI1103, ASI1140, ASI1146, ASI1177, ASI1183, ASI1195, ASI1321, ASI1331, ASI1336 were relatively weakly resistant. The average number of days for 16.5, and that for harvest was 5.7; the average yield was 142.7 g/2-kg bag. Mycelial growth rates at 10, 15, 20, $25^{\circ}C$ were similar, but that at $30^{\circ}C$ was lower. The mushroom yield was highly correlated with the number of days for pinheading and harvest.

• Journal of Mushroom
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• v.13 no.3
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• pp.243-249
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• 2015

Agaricus bitorquis is an edible white mushroom of the genus that is cultivated at high temperature($25{\pm}1^{\circ}C$) unlike A. bisporus is cultivate at $16{\pm}2^{\circ}C$. Unlike Agaricus bisporus, an edible white A. bitorquis mushroom is cultivated at high temperature ($25{\pm}1^{\circ}C$). Most farmers cultivate this mushroom for a long cultivation period in Korea. For this reason, we made heterokayons to develop a new cultivar that generate fruitbodies for short cultivation period. Over one hundred SSIs(single spores isolates) were collected from selected A. bitorquis ASI1151 and ASI1349 strains. Seventy-three SSIs were germinated on CDA(compost dextrose agar) media after 20 days (minimum) or 83 days (maximum) incubation under different media condition. The mycelial growth rate of germinated SSIs was different. 9 homokaryons in ASI 1151 and 11 homokaryons in ASI 1349 from SSIs were selected by OPN-02 primer in RAPD analaysis. Also this primer was used to select heterokaryon that cross among each homokaryon with compatible locus. Therefore 44 compatible matings were confirmed of 99 crossed lines.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.378-385
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• 1994

Interspecific somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus and Pleurotus sajor-caju. The fusion products between incompatible strains did not form clamp connections. Fruiting body of the clampless fusants was induced by light-dark cycle on saw-dust-rice bran substrate in glass bottles. Out of them, seven somatic hybrids produced fruiting bodies of intermediate morphology of the two species. Light and low temperature were the initiating factors for the development of clamped hyphae from the clampless mycelial colonies. All of these basidiocarps had clamp connections. Eight fusants from the six crosses were analysed with the segregation of genetic characters by random spore isolates. In the three combinations, unexpected alleles were shown. Somatic hybrid between P188 (P. ostreatus 2-1 + P. sajor-caju 2-53) and P. florida 2-3 by triple cross produced fruiting bodies similar to those of fusant between P. ostreatus and P. florida. All the genetic charaters from the three strains were shown to segregate and recombine.

• The Korean Journal of Mycology
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• v.41 no.2
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• pp.91-96
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• 2013

Two strains Pleurotus eryngii 'Aeryni' and 'Na' carrying superior traits of a pileus and a earliness of harvest were selected to improve previously bred strains by single crosses. New hybrid, Aeryni 3 (Aeryni10 ${\times}$ Na5) showed superiority to other hybrids in yield, fruit body shape and days for harvest. The new strain, Aeryni 3 was harvested earlier than Keunneutari No. 2 by 2~3 days, and yielded 110.5 g/bottle (850 mL) which was 108% of that of Keunneutari No. 2. The ratio of diameter of pileus and stipe was 1.8 indicating that new stain will be likely low damage rate of fruit body during a distribution, and that was better than 2.1 of Keunneutari No. 2. A sensory test of taste of the new strain showed that 84.7% of evaluation panels selected "very good" while that of Keunneutari No. 2 was 55.5%. In purchasing intent test, 86.9% of panels will buy the new stain whereas 46.8% will buy Keunneutari No. 2 implicating that the new strain will likely be more marketability than previously bred strain.