• Journal of Life Science
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• 제12권2호
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• pp.53-56
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• 2002

The baculovirus expression vector system (BEVS) is one of the powerful heterologous protein expression systems using insect cells. As a result this has become a hot issue in the fleld of biotechnology. The advantage of the BEVS is that the large-scale production of heterologous proteins, which undergo posttranslational modification in the endoplasmic reticulum (ER), can be accomplished. Altrough posttranslational modification of heterologous proteins in insect cells is more similar to mammalian cells than yeast, it is not always identical. Therefore, aggregation and degradation can sometimes occur in the ER. To produce a high level of bioactive heterologous proteins using BEVS in insect cells, the prerequisite is to completely understand the posttranslational conditions that determine how newly synthesized polypeptides are folded and assembling with ER chaperones in the ER lumen. Here, we provide information on current BEVS problems and the possibility of successful heterologous protein production from mammalian cells.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1719-1727
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• 2014

In the present study, whispovirus immediate early 1 promoter (ie-1) was used to initiate surface expression of the hemagglutinin (HA) protein of Egyptian H5N1 avian influenza virus (AIV) by using the baculovirus expression vector system. The HA gene and whispovirus ie-1 promoter sequence were synthesized as a fused expression cassette (ie1-HA) and successfully cloned into the pFastBac-1 transfer vector. The recombinant vector was transformed into DH10Bac competent cells, and the recombinant bacmid was generated via site-specific transposition. The recombinant bacmid was used for transfection of Spodoptera frugiperda (Sf-9) insect cells to construct the recombinant baculovirus and to induce expression of the HA protein of H5N1 AIV. The recombinant glycoprotein expressed in Sf-9 cells showed hemadsorption activity. Hemagglutination activity was also detected in both extra- and intracellular recombinant HAs. Both the HA and hemadsorption activities were inhibited by reference polyclonal anti-H5 sera. Significant expression of the recombinant protein was observed on the surface of infected insect cells by using immunofluorescence. SDS-PAGE analysis of the expressed protein revealed the presence of a visually distinguishable band of ~63 kDa in size, which was absent in the non-infected cell control. Western blot analysis confirmed that the distinct 63 kDa band corresponded to the recombinant HA glycoprotein of H5N1 AIV. This study reports the successful expression of the HA protein of H5N1 AIV. The expressed protein was displayed on the plasma membrane of infected insect cells under the control of whispovirus ie-1 promoter by using the baculovirus expression vector system.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.318-322
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• 1999

A new baculovirus transfer vector (pPGP404) was constructed to increase the expression level of human thrombopoietin (hTPO) in insect cells. In pPGP404, hTPO was cloned next to the AcNPV polyhedrin-gp64 dual promoter and the leader sequence of hTPO was substituted with that of gp64. A recombinant baculovirus, AcPGP404, was constructed by using pPGP404 as a transfer vector. hTPO was expressed in AcPGP404-infected TN5 cells and it was observed that the expression levels of hTPO in TN5 cells increased three-fold ($6.0 {\mu}g/ml^{-1}$) compared to the level expressed under the control of the polyhedrin single promoter. These results indicate that the polyhedrin-gp64 dual promoter system would be useful for expression in large quantities of recombinant proteins in insect cells.

• The Plant Pathology Journal
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• 제23권2호
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• pp.57-61
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• 2007

Whitefly-transmitted geminiviruses (WTGs) are economically important pathogens causing serious damage on tomato and chilli pepper in Indonesia. Geminiviruses are readily transmitted by its insect vector, sweetpotato whitefly (Bemisia tabaci). However, greenhouse whitefly (Trialeurodes vaporariorum), another species of whitefly, is commonly found together with B. tabaci in the field. Incidence of yellow leaf curl disease in tomato and chilli pepper is probably correlated with the population of whitefly complex. It is becoming important to find the role of T. vaporariorum in the spread of the disease. Therefore, research is conducted to study the characteristic relationship between tomato leaf curl begomovirus (ToLCV) and two species of whitefly. The two species of whitefly, B. tabaci and T. vaporariorum, was capable to transmit ToLCV although it was evidenced that B. tabaci is more effective as insect vector of ToLCV in tomato and chilli pepper. A single B. tabaci was able to transmit ToLCV to tomato with a minimum acquisition and inoculation access period of 10 h. Transmission of ToLCV by T. vaporariorum required at least 10 insects per plant with a minimum acquisition and inoculation access period of 24 h. The transmission efficiency will increase with longer acquisition and inoculation access period of the insect and the higher number of insect per plant.

• Mycobiology
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• 제45권3호
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• pp.160-171
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• 2017

Larvae of Bradysia agrestis, an insect vector that transports plant pathogens, were sampled from geographically isolated regions in Korea to identify their cutaneous fungal and bacterial flora. Sampled areas were chosen within the distribution range of B. agrestis; each site was more than 91 km apart to ensure geographical segregation. We isolated 76 microbial (fungi and bacteria) strains (site 1, 29; site 2, 29; site 3, 18 strains) that were identified on the basis of morphological differences. Species identification was molecularly confirmed by determination of universal fungal internal transcribed spacer and bacterial 16S rRNA gene sequences in comparison to sequences in the EzTaxon database and the NCBI GenBank database, and their phylogenetic relationships were determined. The fungal isolates belonged to 2 phyla, 5 classes, and 7 genera; bacterial species belonged to 23 genera and 32 species. Microbial diversity differed significantly among the geographical groups with respect to Margalef's richness (3.9, 3.6, and 4.5), Menhinick's index (2.65, 2.46, and 3.30), Simpson's index (0.06, 0.12, and 0.01), and Shannon's index (2.50, 2.17, and 2.58). Although the microbial genera distribution or diversity values clearly varied among geographical groups, common genera were identified in all groups, including the fungal genus Cladosporium, and the bacterial genera Bacillus and Rhodococcus. According to classic principles of co-evolutionary relationship, these genera might have a closer association with their host insect vector B. agrestis than other genera identified. Some cutaneous bacterial genera (e.g., Pseudomonas) displaying weak interdependency with insect vectors may be hazardous to agricultural environments via mechanical transmission via B. agrestis. This study provides comprehensive information regarding the cutaneous microflora of B. agrestis, which can help in the control of such pests for crop management.

• 한국산림과학회지
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• 제95권5호
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• pp.543-550
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• 2006

Distribution patterns of two pine sawyer species (Monochamus alternatus which is the main vector insect and M. saltuarius which is the potential insect vector of the pine wood nematode) were investigated in Korea. The data were collected at 89 study sites which were chosen to cover the whole region of South Korea. The selected pine trees were killed in early April and left for I year in the pine stands to be egg-laid by the pine sawyers. Emergence of the beetles from the dead pine trees was checked from early April to late July. M. saltuarius was the most abundant in the mid to northern areas of South Korea, whereas M. alternatus in Jeju-do, southernmost island of Korea. Considering temperature distribution patterns in areas where the two species occur, their thermal distribution boundary may be formed around $13.2^{\circ}C$ of annual mean temperature. The hypothesized distribution map of the two Monochamus species under the invasion of pine wilt disease is suggested on the base of thermal distribution of Korean peninsula.

• 농약과학회지
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• 제15권4호
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• pp.490-494
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• 2011

벼줄무늬잎마름병(Rice stripe tenuivirus; RSV)은 벼에 발생하는 주요 바이러스병으로 2000년대 이후 서해안을 중심으로 벼에 발생하여 큰 피해를 주고 있다. RSV는 벼의 주요 해충인 애멸구에 의해 영속적으로 전염되기 때문에 애멸구의 적절한 방제로 피해를 경감시킬 수 있다. RSV의 방제전략을 수립하기 위해서는 RSV와 애멸구 간의 생태학적 연구가 필요하며, 특히 애멸구의 흡즙 및 감염생태가 결정적인 키포인트가 될 수 있다. 따라서 본 연구에서는 RSV에 감염된 벼에서 애멸구의 흡즙 및 감염특성을 조사하였다. 애멸구의 영기별 RSV 전염능력을 조사한 결과, RSV 감염주에서 3일 이상 흡즙한 4령충 이상에서 바이러스 전염능력이 높았다. RSV 이병주를 3일간 흡즙하였을 때 성충의 전염능력은 44.8%인데 비해 4령충과 5령충의 전염능력은 69.2%와 67.9%로 성충보다 4, 5령충의 전염능력이 높았다. 이병주 흡즙시 온도에 따른 보독충의 전염능력 조사 결과에서는 $30^{\circ}C$에서 전염능력이 가장 높았으며, $30^{\circ}C$에서 3일간 흡즙하였을 때 전염능력은 93.3%로 $25^{\circ}C$와 $20^{\circ}C$의 70.6%, 43.8%에 비해 현저히 높았다.

• 셀메드
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• 제4권2호
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• pp.8.1-8.6
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• 2014

Despite the remarkable advancements of the modern medicine, the traditional system of medicine (TSM) still serves as a potential primary health care modality in the in low- and middle-income countries. The recent reports suggest that there is a renewed interest has been observed towards TSM in the developed countries too, because of the adverse side-effect of modern medicines. Medicinal plants have been widely serving as a rich source of therapeutic agent. Ethiopia is one of the most reserves rich countries in the world. It is renowned for well-diversified and natural resources in terms of its unique flora and fauna. Ethiopian deep-rooted tradition and culture largely depends on the usage of plants for their religious ceremonies, impressive festivals, traditional medicinal uses and other basic necessities. The present scrutiny is an attempt to understand the omnipotent nature of an Ethiopian Ethnomedicinal plant called Tinjute [vernacular name (local native language, Amharic); Otostegia integrifolia]. There are several studies suggest that Tinjute can be used as a natural medicine or health-promoting agents for various disorders and ailments. Nevertheless, in Ethiopia, it is renowned as an insect repellent to drive-away insect vector of diseases, particularly mosquitoes in the early evening. However, there are many more issues and challenges which must be urgently addressed to scientifically formulate various potent, efficacious, safe and highly selective phytotherapeutic agents and insects' repellent from the Tinjute plant in the near future.

• International Journal of Industrial Entomology and Biomaterials
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• 제34권1호
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• pp.1-5
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• 2017

Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

• Journal of Forest and Environmental Science
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• 제34권5호
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• pp.395-404
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• 2018

Since the mass mortality of Quercus mongolica has been first reported in Gyeonggi province at 2004, the disease spread rapidly over Korean peninsula annually. Ambrosia beetle (Platypus koryoensis) was known as the insect vector of oak wilt fungus, Raffaelea quercus-mongolicae, and control methods of the disease had mainly been focused on eradication of insect vector. However, for the efficient management of the disease, combined control methods for both of the pathogenic fungus and insect vector are strongly required. As one of the efforts to suppress the pathogenic fungus, antifungal activities of Streptomyces isolated from oak forest soil were assayed in this study. Optimum culture condition for the selected isolates was also studied, As a result, Streptomyces blastmyceticus cultured in PDB (Potato Dextrose Broth) at $25^{\circ}C$ for 1 week showed the strongest antifungal activity against oak wilt fungus. Mycelial growth inhibition rates (MGIRs) of Streptomyces isolates were compared on culture media supplemented with heated and unheated culture filtrates of S. blastmyceticus. MGIRs on culture media with unheated culture filtrates were generally higher than those on culture media with heated culture filtrates. Antagonistic mechanism to get involved in the inhibition of hyphal growth and spore formation of the pathogen is due to the antifungal metabolites produced by Streptomyces. This study will provide the fundamental information in developing biocontrol agents for the environment-friendly management of oak wilt disease.