Most traditional genome sequencing projects involving infectious viruses include culturing and purification of the virus. This can present difficulties as an analysis of multiple populations from multiple locations may be required to acquire sufficient amount of high-quality DNA for sequence analysis. The electrophoretic method provides a strategy whereby the genomic DNA sequences of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) were analyzed by purifying it from host DNA by pulsed-field gel electrophoresis, thus simplifying sampling and labor time. The genomic DNA of infected P. rapae was embedded in agarose plugs, digested with a restriction nuclease and methylase, and pulsed-field gel electrophoresis (PFGE) was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the separated viral DNA. The double-stranded circular genome of PiraGV-K encodes 120 open reading frames (ORFs), covering 92% of the sequenced genome. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (~99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF11), involved in the liquefaction of the host, were found in the genome. The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the pulsed field electrophoretic method. The method appears to be applicable to the analysis of genomes of large viruses. The chitinase, identified by PiraGV-K genome sequence, was functionally characterized by quantitative PCR, Western blot analysis, immunohistochemistry and transmission electron microscopy.

The insect baculovirus expression vector system (BEVS) is useful for the production of biologically active recombinant proteins. However, the overexpression of foreign proteins in this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we have developed a versatile baculovirus expression and secretion system using the Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion improved the secretion and antibacterial activity of recombinant nuecin proteins. Thus, bPDI gene fusion is a useful addition to the BEVS for the large-scale production of bioactive recombinant proteins.

한의학에서 승마(Cimicifuge dahurica, C. dahurica), 황련(Coptis chinensis, C. chinensis), 황백 (Phellodendri amurense, P. amurense) 추출물은 해독작용을 하며 후박(Magnol obovata, M. obovata) 추출 물은 항균 효과가 있다고 알려져 있다. 본 연구에서는 승마, 황련, 황백, 후박 추출물을 혼합하여 기능성 화장품 소재로써의 이용 가능성을 검토하였다. MTT assay를 이용한 세포 독성평가와 DPPH와 ABTS를 이용한 항산 화 효과를 측정한 결과, 단일 추출물보다 추출혼합물이 더 안전함을 확인 하였으며 농도 의존적으로 항산화 활성 이 증가됨을 확인하였다. 뿐만 아니라 NO 생성을 억제함으로써 우수한 항염증 효과가 있을 것으로 예상되며, 섬유아세포에서 UVB로 유도된 세포 손상을 보호하면서 type 1 pro-collagen 합성량이 증가되는 것을 관찰 하였다. 또한 추출 혼합물은 melanin 합성과 tyrosinase 활성 및 melanin 합성 관련 인자 및 효소들(MITF, tyrosinase, TRP-1, TRP-2)의 발현을 효과적으로 억제하였다. 이러한 연구결과로 보아 승마, 황련, 황백, 후박 추출 혼합물은 항노화 및 미백 등 기능성 화장품 소재로써 개발 가능성이 높을 것으로 사료된다.

히솝(Hyssopus officinalis)은 통화식물목 꿀풀과의 여러해살이풀로 방부제, 기침, 거담제 등 일반적으 로 방향족 허브와 약용식물로 사용되어왔다. 본 연구에서는 항산화 및 미백소재의 개발을 위해 히솝을 열수와 주정으로 추출하여 각 추출물의 in vitro 상에서의 항산화 효과 및 B16F10 melanoma 세포에서 멜라닌 생성 저해효과를 평가하였다. DPPH와 ABTS assay 시험법을 이용하여 항산화 효과를 측정한 결과, 히솝 추출물의 항산화 활성이 모두 농도 의존적으로 증가하였다. 또한 히솝 추출물은 tyrosianse 활성을 저해시켰으며, B16F10 세포에서 UVB로 증가된 활성산소와 α-MSH로 유도된 멜라닌 또한 감소시키는 효과를 보였다. 멜라 닌 생합성에 관여하는 유전자의 발현에 미치는 영향을 알아보기 위해 RT-PCR을 실시한 결과 히솝 추출물에 의해 MITF, tyrosinase, TRP-2의 발현이 감소되는 것을 확인할 수 있었다. 이러한 결과에 따라 히솝 추출물은 미백개선을 위한 화장품 소재로서 이용 가능성이 높을 것으로 사료된다.

Carotenoids are vital pigments responsible for yellow, orange and red color in plants. In Capsicum, capsanthin-capsorubin synthase (CCS), phytoene synthase (PSY), β-Carotene hydroxylase (CRTZ-2) and lycopene β-cyclase (LCYB) were identified to be involved in the carotenoids synthesis pathway. Previously molecular markers based on the CCS and PSY genes have been developed to distinguish fruit colors in pepper. However these markers can distinguish fruit colors of limited pepper genotypes. Therefore, there is need of developing additional markers for accurate prediction of fruit colors using molecular markers. In this study carotenoids contents of 16 pepper accessions were analyzed and the CCS, PSY, CRTZ-2, LCYB genes were sequenced to identify the genes affecting the fruit color. Among all the analyzed carotenoids, capsanthin was accumulated in much higher amount in red and orange fruits (1100-2500 mAU·min and 30-500 mAU·min respectively) while violaxanthin (20-1200 mAU·min) was accumulated more in yellow fruits. Sequence analysis revealed that deletions and two frame shift mutations in CCS gene for yellow accessions. Frame shift mutations of the PSY gene were detected in two orange accessions. These results show that mutations in CCS and PSY genes affect the fruit colors of pepper, and markers can be developed using mutations of these genes.

Herb extracts commercially used in Korea were screened for PPAR-γ agonist test and α-glucosidase inhibition assay. Total 16 herb plants had a PPAR-γ agonist activity. Specially, Alisma orientale Juz (108.41%), Ephedra sinica (98.22%), Sasa japonica Makino var. purpurascens Nakai (140.68%), Astragalus membranaceus Bunge (106.79%) and Cnidium officinale Makino (113.00%) showed high PPAR-γ agonist activity rate compared with rosiglitazone's (167.46%). And Cornus officinalis S. et Z. (90.3%), Cinnamomum cassia Blume (89.2%), Psoralea corylifolia L. (89.8%), Paeonia japonica (Makino) Miyabe (92.4%) and Paeonia suffruticosa Andr (93.2%), showed high α-glucosidase inhibition rates. These results support previous reports of the efficacy of Oriental medicinal plants used for diabetes mellitus.

Muscle strength and endurance activities of Korean ginseng (Panax ginseng C. A. Meyer; KG) were compared with those of wild simulated cultivation ginseng (WCG) in mice. Fifty male ICR mice were divided into five groups: A (vehicle); B (WCG 100 mg/kg); C (WCG 500 mg/kg); D (KG 100 mg/kg); E (KG 500 mg/kg). Subsequently, the mice were subjected to the forced swimming test (FST) and treadmill test at the 4th and 7th weeks. The glycogen content in the muscle and blood analysis (levels of glucose, triglyceride (TG), IGF-1) were also performed immediately after the last FST and treadmill test at the 7th week. Immobility times in FST were shorter in WCG- than KG-treated groups, and the results of the treadmill tests were also significant except for KG-treated at 100 mg/kg. The glycogen content was increased in both groups with a peak at 500 mg/kg of WCG groups. Serum concentrations of TG and glucose were decreased by administration of KG and WCG and all treated groups showed increase in the level of IGF-1 in serum. These results suggest that KG and WCG supplementations are effective in escalating the muscle strength and endurance.

The groundnut or cultivated peanut (Arachis hypogaea L.) in Korea consists of 36 domestic varieties which have been developed and registered as cultivars for the public during last 25 years. To screen and identify of Korean peanut varieties and genetic resources, we present a simple and reliable method. A methodology based on simple sequence repeat (SSR) markers developed and widely used for prominent gene identification and variety discrimination. For identification of those 36 Korean peanut varieties, 238 unique peanut SSR markers were selected from some previously reported results, synthesized and used for polymerase chain reaction (PCR). Data were taken through acryl amide gel electrophoresis and changed into proper formats for application of data mining analysis using Biomine (all-in-one functional genomics data mining program). Consequently, twelve SSR primers were investigated and revealed the differences between those 36 varieties. These primer pairs amplified 27 alleles with an average of 2.3 allele per primer pair. In addition, those results showed genetic relationship by classification method within 36 varieties. The approach described here could be applied to monitoring of our varieties and adapting to peanut breeding program.