Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.

Aujeszky's disease (AD), also called pseudorabies, is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Aujeszky's disease virus (ADV) virions contain several envelope glycoproteins. Among them, glycoproteins gB, gC and gD are regarded as the major immunogenicity proteins and the antibodies induced by them can neutralize virus in vitro or in vivo. In this study, we investigated expression of these glycoproteins using the bacterial and baculovirus expressionn system. Successful expression of ADV glycoproteins in E. coli was confirmed by SDS-PAGE and Western blot analysis and their optimal expression condition was determined. However, the recombinant proteins generated in the bacterial expression system which lacks glycosylation process frequently lose their biological activity. We tried to express the ADV glycoproteins using the baculovirus expression vector system. The recombinant gB, gC and gD were detected at approximately 100, 60 and 50 kDa on SDS-PAGE and Western blotting, respectively. The optimal expression conditions were determined for MOI(multiplicity of infection) and post-infection days. One MOI and 4 or 5 days post-infection were the best conditions for the expression of the ADV glycoproteins in Sf21 cells. We are currently investigating the antigenicity of recombinant proteins using experimental animals.

The four genetically distinct isolates have been identified previously from Bombyx mori nucleopolyhedroviruses (BmNPVs) isolated in Korea. To further understand the complex of viruses infecting Bombyx mori, the genome of BmNPV-K1 and K4 strains was completely sequenced and analyzed in comparison with the genome of other sequenced baculoviruses including previously reported BmNPV. BmNPV-K1 consisted of 127,542 bp and 133 open reading frames (ORFs) of 150 nucleotides or longer with minimal overlap have been identified. In contrast, BmNPV-K4 consisted of 128,615 bp and 134 open reading frames (ORFs). Although gene arrangement is virtually identical, the genome of BmNPV-K4 is 1,073 bp longer than BmNPV-K1. This was related to the more existence of bro genes in BmNPV-K4. To investigate the relationship between BmNPV-K1 and K4, phylogenetic analysis with each member of the paired ORFs was performed. The sequence data suggest that BmNPVK1 and BmNPV-K4 are closely related but have diverged and evolved into two separate strains. This was study to identify highly related but separately evolving viruses in the same insect host and geographic location. We are currently comparing the differences of these BmNPV genomes to elucidate characteristics of each virus.

Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). To enhance the expression level of baculovirus vector system, we constructed several fusion vectors using various fragments of the polyhedrin. The polyhedrin fragments were genetically fused to the enhanced green fluorescent protein (eGFP) under the control of polyhedrin promoter, and their expressions were analyzed in Sf21 insect cells. Expression of the fusion protein was identified by SDS-PAGE and Western blot analysis using anti-GFP and anti-Polyhedrin. The expression level of eGFP was markedly increased by the fusion of partial polyhedrin. Also, the fluorescence intensity of fusion proteins was higher than that of non-fusion protein. Confocal laser scanning microscopy demonstrated that fusion proteins were localized to the cytosol or nucleus of insect cells. In additional, the glycoprotein E2 (gE2) of classical swine fever virus (CSFV) expressed by the these vectors was dramatically increased and its immunogenicity was proofed using experimental animal guinea pigs that were immunized with the partial polyhedrin containing gE2. This study provides a new option for the higher expression of useful foreign recombinant protein by using the partial polyhedrin in BEVS.

Hereditary dentin defects consists of dentin dysplasia(DD) and denti nogenesis imperfecta(Dr) ‘ The Dl associated with osteogenesis imperfecta has been classified as DI type 1. whereas isolated inherited defects have been categori zed as DI types II and III , However‘ whether DI type III should be considered a distinct phenotype 01' a variation of DI type 1I is debatable , Recent genetic findings have focused attention on the role of the dentin sialo phosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin, We have identified novel mlltation( c,727G - > A, p,D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth These findings sllggest that DI type II and III are not separate diseases bllt rather the phenotypic variation 01' a s ingle disease

Stevia rebaudiana (Asteraceae), a perennial plant, has been used as a low-calorie sweetener and is being developed as a therapeutic agent for diabetes, hypertension, myocardial diseases, and microbial infections. Despite the common use of its leaves and stem, the bioavailability of the components present in S. rebaudiana flowers, when used as ingredients of cosmetics, has not been well investigated. Herein, we investigated the antioxidative and antimelanogenic effects of an aqueous extract of S. rebaudiana flowers (Stevia-F). Total flavonoid and phenolic content in Stevia-F were determined to be 8.64 ± 0.23 ㎎ of quercetin equivalents/100 g and 631.5 ± 2.01 ㎎ of gallic acid equivalents/100 g, respectively. The IC50 values of Stevia-F for reducing power, and 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical, hydrogen peroxide, and nitric oxide scavenging activities were 5541.96, 131.39, 466.34, and 10.44 ㎍/mL, respectively. Stevia-F showed inhibitory effects on the tyrosinase (IC50 = 134.74 ㎍/mL) and α-glucosidase (IC50 = 114.81 ㎍/mL) activities. No significant cytotoxicity of Stevia-F was observed in B16F10 cells, treated with up to 100 ㎍/mL of the extract for 24 and 48 h (p > 0.05). Stevia-F (1–100 ㎍/mL) suppressed α-melanocyte stimulating hormone-induced melanin production in B16F10 cells (p < 0.05) and also inhibited the cellular tyrosinase activity (p < 0.05). Overall, our results show that Stevia-F possesses potential for inhibiting tyrosinase and α-glucosidase activities and has significant antioxidant capacity. The antimelanogenic potential of Stevia-F should extend the usage of S. rebaudiana flowers in the development of skinwhitening products.

Angelica tenuissima, also known as Ligusticum tenuissimum, is classified as a food-related plant and has been used as traditional medicines treating headache and anemia in Asia. However, its anti-melanogenic effect has not been reported in detail. When the extract of Angelica tenuissima (ATE) was prepared by the extraction with 70% EtOH at 80°C (final yield = 22%), the contents of decursin and Z-ligustilide in ATE were determined 0.06% and 8.43%, respectively. Total flavonoid and phenolic content in mg ATE were 5.52±0.07 ㎍ quercetin equivalents and 237.27±13.24 ㎍ gallic acid equivalents, respectively. Antioxidant capacity of ATE determined by DPPH and ABTS assay was increased with a dose dependent manner up to 1000 ㎍/㎖. The amount of melanin synthesis followed by α-melanocyte stimulating hormone on B16F10 cells were significantly reduced in the presence of ATE (250 to 1000 ㎍/㎖, p<0.05). ATE (125 to 1000 ㎍/㎖, p<0.05) suppressed the tyrosinase activity but did not show any significant effect on α-glucosidase activity at the same condition. Taken together, ATE possesses tyrosinase inhibitory potential with significant antioxidant capacities. These effects of ATE might be involved in suppression of melanin synthesis, at least, in B16F10 cells. The anti-melanogenic potential of ATE will provide an insight into developing a new skin whitening product.

EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

Backgoound : This study was conducted to serve as a basis for the selection of superior lines by analyzing the content of antioxidant component and antioxidant activity in Schisandra chinensis Collections Methods and Results : In order to examine antioxidant component and antioxidant activity, 154 species of Schisandra chinensis from whole country were used. Antioxidant component was investigated by total flavonoid content and total phenolic content and antioxidant activity was evaluated by DPPH radical scavenging activity and ABTS radical scavenging activity. As a result, the total amounts of flavonoids was highest in SC-20 with 5.03 ㎎/g. However the content of polyphenols showed highest in SC-22 with 2.76 ㎎/g. In addition antioxidant activity results were also relatively high in SC-22. The IC50 value was 548 ㎍/㎖ in DPPH radical scavenging and 640 ㎍/㎖. in ABTS+ radical scavenging. Conclusion : As demand for high income crop has increased, new cultivar breeding is required to produce high quality Schisandra chinensis From this study, analyses of antioxidant component and antioxidant activity in collection can be used for new Schisandra chinensis breeding. Especially SC-22 can be superior lines with high antioxidant component and antioxidant activity.

Background : Stable ginseng production is highly dependent on specific climate conditions. Recently, ginseng yield and quality are negatively affected by climate changes, particularly global warming. Thus, it is imperative to apply a new systematic cultivation method and to develop new varieties with enhanced heat resistance to cope with elevated temperatures. Up to date, ginseng breeding program has mainly focused on the quantity and process ability of red ginseng. New varieties with increased resistance to diverse abiotic stresses need to be developed. Methods and Results : In this study, 13 varieties and 100 germplasms were screened for resistance to heat stress. To measure heat resistance, seedlings were transplanted to a pot 3 times in threes. Two months later, the pot was placed in a temperature chamber at 46℃ for 1 hour, a critical condition that ginseng appears to stop cellular respiration. After waiting for 1 day, the pot was examined for survival rate and fluorescent reaction. Fluorescent reaction was tested according to Nedbal et al. (2000) and Park et al. (2010) and with an image fluorometer. After testing chlorophyll fluorescent reaction, Fm/Fo, Fv/Fm and Rfd were used as screening indices for high temperature resistance. It was found that the adequate values for Fm/Fo, Fv/Fm and Rfd were over 2.2, over 0.55 and over 0.8, respectively. Also, the degree of the above-ground damage is investigated in the 4-year old field. Under our heat stress conditions, susceptible varieties (Chunpoong) and germplasm lines showed drooping and wilting leaf phenotypes, whereas the leaves of a resistant line (named Eumseong 11) remained healthy. Eumseong 11 had the highest values with 2.3, 0.61 and 0.98 in fluorescent test. In addition, compared to the susceptible lines which were wilted in 4-year old field, no detrimental phenotypes were observed in Eumseong 11. Conclusion : ‘Eumseong11’ was selected as a superior line with increased resistance to heat stress. We are now testing how Eumseong11 responds to other abiotic stresses. Our effort will contribute to increase the farmers' income.

Background : Panax ginseng C. A. Meyer is a slow-growing perennial herb that is cultivated in shading condition. Climate change occur around the world that make a lot of problem such as damage of high temperature, drought, salinity and disease. The problems lower the ginseng productivity that cause income reduction of farmers. To achieve stable ginseng production, development of elite varieities resistant to abiotic and biotic stresses is consistently required. It is very time consuming process in order to develop new ginseng varieties because ginseng flowers after 3 years of growth. So, early selection system of elite line must be established. This study was conducted to develope efficient ginseng breeding techniques for early identification of heat or salinity resistance. Methods and Results : Ginseng petioles was soaked in mixed salts solution consisting of KNO3, KH2PO4, MgSO4․H2O in order to test resistant or susceptible salinity. The degree of resistance was quantified according to damage size. Also, ginseng lines transplanted in pot were treated 46℃ for 1 hour and then chlorophyll fluorescence reaction were measured in order to test resistant or susceptible high-temperature. The measured values such as Fm/Fo, Fv/Fm, Rfd were differentiated between resistant and susceptible line. Conclusion : Several lines showed that they are resistance to high temperature or salinity. The selected lines will be utilized for parents to develop new varieties.

Background : Management of air temperature are known to primarily affecting on physiological properties and yield in plant. Methods and Results : The effect of air temperature on characteristics of photosynthesis and chlorophyll fluorescence in Schisandra chinensis Baillon were investigated under controlled temperature using growth chamber. Net photosyntheis rate, transpiration was measured at 1,000 μmol m-2 s-1 of photon flux density and chlorophyll fluorescence was analyzed by OJIP method. Net photosyntheis rate and transpiration rate was higher in treatment of 25℃. As results of chlorophyll fluorescence by OJIP analysis, maximum quantum yield (Fv/Fm) of photosystem II (PSII) and PIabs was higher in treatment of 25℃ which reflects the relative reduction state of PSII. But in treatment of 35℃ the relative activities per reaction center such as ABS/RC, DIo/RC were higher than in treatment of 25℃ which implied that the relative reduction of electron transport at PSI and increasement of photo inhibition at reaction center. Conclusion : This result implies that 25℃ of air temperature may be a adequate temperature to improving the efficiency of photosynthesis through controlling a photosystem in Schisandra chinensis Baillon.

Background : When ginseng seeds were gathered, the seeds were unripe. To grow immature embryo definitely, special treatment called dehiscence must be performed. Even though dehiscence is completed, most ginseng seeds are on enforced dormancy. The breaking seed dormancy is generally achieved using cold treatment. Also it is reported that gibberellin treatment can replace the treatment. It is very time consuming process in order to develop new ginseng cultivar because ginseng flowers after 3 years of growth. To shorten the ginseng breeding period, it is necessary to establish fast generation progress. Therefore, this study examined the possibility of breaking seed dormancy of ginseng using GA3 treatment and alternating temperature. Methods and Results : Seeds were obtained from local variety fruit which is not inbred. Gibberellin of 100 ppm was treated at seeds for 24 hours. Fixed cold condition was treated on both –2℃ and 2℃. Alternating cold condition was treated on 2℃ and then –2℃, finally 2℃. Fixed and alternating temperature was continued for 15, 30, 45, 60, 90 days that 15 days of alternating temperature is first 2℃ for 5days and then -2℃ for 5days, finally 2℃ for 5days. The other treatment periods such as 30, 45, 60, 90 days mean 10, 15, 20, 30 days respectively. Each of 48 seeds were sowed on tray in greenhouse at 3 replication. Experimental plot was completely randomized. Conclusion : Seeds untreated with GA3 were germinated little and there is no difference between 2℃ and –2℃. Alternating temperature until 60days made no difference with fixed temperature but germination rate increased up to 70.8% when seeds were treated for 90days. Germination of seeds treated with GA3 is much higher than untreated seeds especially combined with alternating temperature.

Measurements of closely related sets of classical and truss dimensions were analyzed to discriminate species of scorpaenidae including the dark banded rockfish, Sebastes inermis, the black rockfish, S. schlegeli, and gobioninae including the striped shiner, Pungtungia herzi, and the slender shiner, Pseudopungtungia tenuicorpa. The measurements of the dimensions were arc sin square root transformed, and compared as a function of the standard length of each species for statistical analysis. For values of the classical dimensions of the rockfish, 6 were greater for the dark banded rockfish than for the black rockfish, 1 value was smaller for the former, and for 2 values there was no statistically significant difference (P > 0.05). For values of the classical dimensions of the shiners, 9 values were greater for the striped shiner than for the slender shiner, 2 values were smaller for the former, and for 1 value there was no statistically significant difference (P > 0.01). For values of the truss dimensions of the rockfish, 6 were greater for the dark banded rockfish than for the black rockfish, 1 was smaller for the former, and for 4 values there was no statistically significant difference (P > 0.05). For values of the truss dimensions of the shiners, 13 values were greater for the striped shiner than for the slender shiner, 3 values were smaller for the former, and for 6 values there was no statistically significant difference (P > 0.01). The dimension sets used in this study may be useful as taxonomic indicators for discriminating among fish species in Korea.

Although much effort has been made to find agronomically important loci in the soybean plant, extensive linkage disequilibrium and genome duplication have limited efficient genome-wide linkage analyses that can identify important regulatory genes. In this respect, recombination block-based analysis of cultivated plant genomes is a potential critical step for molecular breeding and target locus screening. We propose a new three-step method of detecting recombination blocks and comparative genomics of bred cultivars. It utilizes typical reshuffling features of their genomes, which have been generated by the recombination processes of breeding ancestral genomes. To begin with, mutations were detected by comparing genomes to a reference genome. Next, sequence blocks were examined for likenesses and difference with respect to the reference genome. The boundaries between the blocks were taken as recombination sites. All recombination sites found in the cultivar set were used to split the genomes, and the resulting sequence fragments were named as core recombination blocks (CRBs). Finally, the genomes were compared at the CRB level, instead of at the sequence level. In the genomes of the five Korean soybean cultivars used, the CRB-based comparative genomics method produced long and distinct CRBs that are as large as 22.9 Mb. We also demonstrated efficiency in detecting functionally useful target loci by using indel markers, each of which represents a CRB. We further showed that the CRB method is generally applicable to both monocot and dicot crops, by analyzing publicly available genomes of 31 soybeans and 23 rice accessions.

We examined the morphometric characteristics and fluctuating asymmetry of diploid and triploid marine medaka, Oryzias dancena. We used morphometric parameters the truss and classical dimensions. Significant differences in all the classical and truss dimensions of the diploid and triploid fish. All the dimensions of the triploid fish were greater than those of the diploid fish. The triploid marine medaka shows sexual dimorphism in these characters, and the sexual dimorphism of the triploid marine medaka is similar to that of the diploid marine medaka. Thus, the growth of triploid marine medaka is faster than that of the diploid fish, and it displays clear sexual dimorphism, with male fish having longer dorsal and anal fins than female fish. we examined fluctuating asymmetry of eye diameter, maxilla length, operculum length, number of pectoral fin ray and number of pelvic fin ray. In all experimental groups, Eye diameter and maxilla length showed no significant difference between left side and right side (P>0.05). Trends of operculum length in triploid male group and pectoral fin ray's number in diploid male group showed similar trend with trends of operculum length in triploid female group and pectoral fin ray’s number in diploid female group. However, trends of operculum length in diploid male group and pectoral fin ray's number in triploid male group showed opposite trend with trends of operculum length in diploid female group and pectoral fin ray’s number in triploid female group. Trend of pelvic fin ray's number in all groups showed similar trend with trend of pectoral fin ray’s number in all groups.

Discovery, identification, and informatics of low molecular weight peptide are extensively rising in the field of proteomics research. In this study, we analyzed protein profiles to discover peptide based biomarker for twelve different soybean seeds with three different agronomic types using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). For optimization of SELDI-TOF MS in soybean seed proteome analysis, four different extraction buffers were tested with urea solubilization buffer, thiourea/urea solubilization buffer, phenol extraction buffer, and modified trichloroacetic acid (TCA)/acetone precipitation/urea solubilization extraction buffer. Two different type of ProteinChip arrays, cation exchange (CM10) and anion exchange (Q10), applied to profile peptides. Among the four different extraction buffers, phenol extraction was selected to protein extraction methodology. Numbers of detected peak cluster in twelve soybean seeds were 125 at CM10 and 90 at Q10 array in the mass range from 2 to 40 kDa. Among them, 82 peak clusters at CM10 and 33 peak clusters at Q10 array showed significantly different peak clusters at p<0.00004 (CM10) and p<0.00005 (Q10) among twelve different soybean cultivars. Moreover, 29 peak clusters at CM10 and 17 peak clusters at Q10 array were detected in all cultivars as an ‘universally existed peptide’. In comparison with three different agronomic types, total of 55 peak clusters (CM10) and 23 peak clusters (Q10) were significantly different peak clusters at p<0.00004 and p<0.0001, respectively. In these probability levels, soybean seeds were well discriminated into different cultivar and different type with each other. Also we could find several specific peptide biomarkers for agronomic type.