Classical swine fever virus (CSFV) envelope glycoprotein E2 is the main target for inducing neutralizing antibodies and protective immunity in swine. Here, we report a novel strategy forthe large-scale production of a CSFV E2 subunit vaccine that demonstrates a high immunogenic capability in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (CSFV E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed an approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Mice that were immunized with the granule form of recombinant polyhedra or the soluble form of the fusion protein elicited CSFV E2 antibodies, which indicated that the recombinant polyhedra carrying CSFV E2ΔC were immunogenic. The virus neutralization test showed that the serum from mice that were treated with recombinant polyhedra or the soluble form of the fusion protein contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen and that the recombinant polyhedra containing CSFV E2ΔC as a granule antigen can be used as a potential subunit vaccine against CSFV.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP4, GP5, and M. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the GP4, GP5, and M with SUMO (Small ubiquitin-related modifier), and expressed the fused gene in Bm5 cells and silkworm larvae. Expression of the proteins were analyzed by 12% SDS-PAGE and western blotting using 6xHis tag and porcine anti-PRRSV antibodies. In results, SUMO fused proteins were expressed at a high level in Bm5 cells. The levels of protein using the silkworm larvae is higher than that using Bm5 cells. The fused protein was purified by Ni-NTA affinity chromatography. This study demonstrated that SUMO, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

Bee venom contains a variety of peptides and enzymes, including serine proteases. Here we describe the molecular cloning and characterization of a serine protease (Bt-VSP) isolated from the venom of the bumblebee Bombus terrestris. The Bt-VSP gene consists of six exons encoding a 358-amino acid protein. The form of Bt-VSP detected in bee venom was the 34-kDa mature protein, which is created by cleavage of the catalytic domain of Bt-proVSP between Arg111 and Val112. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. The finding that Bt-VSP acts as a fibrin(ogen)olytic enzyme is similar to a previous finding that Bi-VSP, a venom serine protease of B. ignitus, exhibits fibrin(ogen)olytic activity. We also compared major venom components in honeybee and bumblebee, and found that bumblebee venom contains a larger amount of serine protease. Furthermore, unlike bumblebee venom, which exhibits fibrin(ogen)olytic activity owing to the presence of a serine protease, it is likely that honeybee venom lacks fibrin(ogen)olytic activity.

The Classical Swine Fever Virus (CSFV) is a member of the Pestivirus genus of the Flaviviridae. The genome of CSFV is a positive single-stranded RNA molecule 12.3 kb and contains a single large open reading frame (ORF). The polyprotein composed of eight nonstructural and four structural proteins (nucleocapsid protein C and three envelope glycoprotein E0, E1 and E2). E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. To determine the characteristics of the CSFV, LOM strain, we investigated the nucleotide sequence of the glycoprotein E0, E1 and E2. Comparison of the LOM with the other strains revealed nucleotide sequence identity ranging from 97 to 98%. Expression of the glycoprotein E2 was identified by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies in Sf21 cells. The expression levels of glycoprotein E2 were observed from day 3 and 5 days maximum. In addition, its expression efficiency by media and cell line was investigated. The result showed that High-Five cells and Grace’s insect media for Sf21 were the best conditions for the expression of the glycoprotein E2.

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. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.

We cloned and characterized two peroxiredoxins (Prxs), BiPrx1 (a 1-Cys Prx) and BiTPx1 (a 2-Cys Prx) from the bumblebee Bombus ignitus. The BiPrx1 gene consists of 5 exons, encoding 220 amino acid residues with one conserved cysteine residue. The BiTPx1 gene consists of three exons, encoding 195 amino acid residues with 2 conserved cysteine residues. Recombinant BiPrx1 (27 kDa) and BiTPx1 (25 kDa), expressed in baculovirus-infected insect Sf9 cells, reduced H2O2 in the presence of electrons donated by dithiothreitol. Unlike BiTPx1, however, BiPrx1 did not show reduction activity when thioredoxin was used as the electron donor. Both BiPrx1 and BiTPx1 protected super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Tissue distribution analyses showed the presence of BiPrx1 and BiTPx1 in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting that BiPrx1 and BiTPx1 are not secretable. When H2O2 was injected into B. ignitus bees, BiPrx1 and BiTPx1 transcripts were acutely up-regulated in the fat body tissues. We also demonstrated regulation of BiPrx1 and BiTPx1 expression via reduction of transcript levels in the fat body with RNA interference (RNAi). Under H2O2 overload, the RNAi-induced BiPrx1 knock-down B. ignitus worker bees showed up-regulated expression of BiTPx1. Reciprocally, BiTPx1 RNAi knockdowns showed up-regulated BiPrx1 expression in the fat body. These results indicate that loss of expression of BiPrx1 or BiTPx1 is compensated by up-regulation of expression of the other peroxidase in response to H2O2 overload.

To determine the characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), CA, which was isolated from the serum of an infected pig in 2006, we investigated the nucleotide sequence and expression of the structural ORFs (ORFs 2 to 7) using the bApGOZA system. We found that the structural ORFs 2 to 7 of CA consisted of 3188 nucleotides that were the same as those formed from VR-2332. Comparison of the CA with the other strains revealed nucleotide sequence identity ranging from 89.8 to 99.5%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The CA strain was closely related to the other North American genotype strains but formed a distinct branch with high bootstrap support. Additionally, expression levels of the PRRSV proteins in Sf21 cells were strong or partially weak. The results of this study have implications for both the taxonomy of PRRSV and vaccine development.

The porcine reproductive and respiratory syndrome virus (PRRSV) has six structural proteins which encoded by ORFs 2 to 7 are designated as GP2, 3, 4, 5, M and N, repectively. In this study, we determined the expression of each protein using novel transfer vector, pBmKSK4 which has the polyhedrin promoter of BmNPV and 6xHis tag. The recombinant transfer vector was co-transfected into Bm5 cells along with bBpGOZA DNA. Recombinant virus was purified by plaque assay and amplified in Bm5 cells. Expression of each protein was identified by SDS-PAGE and Western blot analysis using anti-6xHis monoclonal antibody. The expression levels of the structural proteins in Bm5 cells were stronger than the expression system using pBacPAK9 transfer vector in Sf21 cells. As expected, GP5 was expressed at low levels from its structural properties and its toxicity for cells. In addition, each recombinant protein was purified using Ni-NTA spin columns. The ability to produce each protein in the baculovirus system indicates that these could be major candidates for the development of a vaccine against PRRSV.

Background : Bitter gourd (Momordica charantia) is a vegetable with pantropical distribution and contains the active ingredient, such as momordicine and charantin. Moreover, bitter gourd has been reported to exhibit antidiabetic, anticancer, cardiovascular-protective and antioxidant effects. But, the distinctive bitter taste of bitter gourd was not suitable for food preference. This study was conducted to evaluate the improvement effects of bitter taste of bitter gourd with natural fermentation and roasting process. Methods and Results : Bitter gourd fruits were obtained from Headeulnyeokae Co., Ltd.(Gang-Jin 59253, Repubilc Korea). Fruits of bitter gourd were prepared by natural fermentation process for 4h at 25℃. After natural fermentation, the fruits were dried in hot-air dryer for 15h at 49-55℃. The dried fruits were roasted in 500 g batches at 120℃ for 10 min in the electric rotisserie oven. After roasting, fruit pieces were extracted with hot water and cold water. One of them was cool-type, and the other was hot-type bitter gourd tea. In the sensory evaluation, hot tea and cold tea showed the high scores on color, flavor and overall acceptability. Conclusion : These mean that roasted bitter gourd had less bitter, and it could be utilized widely as drink and food material.

This study was carried out to compare the physicochemical characteristics of vinegars fermented from cereal crops such as glutinous and nonglutinous foxtail millet, proso millet, sorghum and adlay, and Incalgyun. The crude protein, minerals and P2O5 contents of vinegars fermented from cereal crops and Incalgyun were higher than circulation brown rice vinegar. Brix degree, turbidity and pH of vinegar fermented from cereal crops and Incalgyun were higher, and total acidity was lower than circulation brown rice vinegar. The glucose content of circulation brown rice vinegar was 4.89 mg/mL, and vinegars fermented from cereal crops were 0.00-5.62 mg/mL. The total organic acid content of circulation brown rice vinegar was 41.92 mg/mL, and vinegars fermented from cereal crops were 12.14-42.31 mg/mL. The major organic acids were acetic acid, succinic acid and citric acid. The total polyphenol content of circulation brown rice vinegar was 0.023 mg/mL, vinegars fermented from cereal crops were 0.286-0.413 mg/mL, and vinegars fermented from cereal crops and Incalgyun were 0.266-0.396 mg/mL. The ABTS and DPPH radical scavenging activities of vinegars fermented from cereal crops were higher than circulation brown rice vinegar.

“Yeonganbyeo”, a new japonica rice variety (Oryza sativa L.), is a mid- maturing ecotype with high lysine content in kernels that was developed by the rice breeding team of National Yeongnam Agricultural Experiment Station (NYAES) in 2001 and released in 2002. This variety was originated from the single cross of Milyang 122/YR13616 Acp1 (in 1992/1993 winter) and was selected by means of a mixed method of bulk and pedigree breeding. The pedigree of “Yeonganbyeo” was YR15815-B-B-B-30 and designated in 1998 as “Milyang 164”. It has about 83cm in culm length with lodging tolerance. This variety is resistant to bacterial leaf blight (K1, K2, K3), stripe virus, and moderately resistant to leaf blast disease. Milled rice kernels of “Yeonganbyeo” has high lysine content of 4.31% (ratio of amino acid components in total protein), a clear translucent with non-glutinous endosperm and clear in chalkness and good at eating quality by pannel test. The yield potential of “Yeonganbyeo” in milled rice is about 5.45 MT/ha at ordinary fertilizer level of local adaptability test. This cultivar would be adaptable to the southern plain of Korea.

A new malting barley variety, Hopum, was developed from the cross between Sacheon 6 and Misato Golden at thenated as Milyang114. It showed good agronomic performance in the regional adaptation yield trials (RYT) from 2001 to 2003 andwas released with the

Suwon295 at the Honam Agricultural Research Institute (HARI) in 2005. An elite line, 953017-BG-BN-BN-53N, was selected in2000 and designated as Milyang126. It showed good agronomic performance in the regional adaptation yield trials (RYT) from2003 to 2005

A new rice variety Manweolbyeo is a japonica type developed by the rice breeding team of Yeongnam Agriculturalivationcondition from the progenies of a single cross of Milyang120/Hwayeongbyeo. The selected line(YR16326-8-3-2) from the prog-enies of it cros

Effects of ambient and elevated ~textrmCO2 and high temperature, and their interactions with zero and applied nitrogen supply (NN-no nitrogen and AN-applied nitrogen) were studied on soybean (Glycine max L.) in 2001. In this experiment, elevated ~textrmCO2 (650 ~mu~textrmmol.~textrmmol-1 ) and temperature (+5~circ ) increased total dry mass at final harvest by 125% and 119% and seed weight per plant by 57% and 105% for NN and AN plants, respectively. Although the influence of temperature and temperature x ~textrmCO2 were not significant, the influences of ~textrmCO2 concentration and temperature x ~textrmCO2 concentration were significant on total dry weight and seed weight, respectively. In particular, seed weight per plant was increased, while weight per one hundred seed weight was decreased with elevated ~textrmCO2 and temperature. The N supply increased biomass and seed weight per soybean plants. The results of this study suggest that the long-term adaptation of soybean growth at an elevated ~textrmCO2 concentration and high temperature might potentially result in a increase in dry matter production and yield.