• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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• pp.126.1-126.1
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• 2007

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• Journal of Microbiology and Biotechnology
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• 제9권2호
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• pp.227-229
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• 1999

Animal cell culture media are usually supplemented with fetal bovine serum (FBS); however, the use of FBS presents certain problems including high cost. By using an adaptation process and the addition of silkworm hemolymph, the FBS concentration can be reduced without causing a significant decrease in cell growth.

• BMB Reports
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• 제43권8호
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• pp.573-578
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• 2010

Several organophosphorus (OP) insecticides can selectively kill the silkworm maggot, Exorista sorbillans (Es) (Diptera: Tachinidae), while not obviously affecting the host (Bombyx mori) larvae, but the mechanism is not yet clear. In this study, the cDNA encoding an acetylcholinesterase (AChE) from the field Es was isolated. One point mutation (Gly353Ala) was identified. The Es-353G AChE and Es-353A AChE were expressed in baculovirus-insect cell system, respectively. The inhibition results showed that for eserine and Chlorpyrifos, Es-353A AChE was significantly less sensitive than Es-353G AChE. Meanwhile, comparison of the I50 values of eserine, dichlorvos, Chlorpyrifos and omethoate of recombinant Es AChEs with its host (Bombyx mori) AChEs indicated that, both Es AChEs are more sensitive than B. mori AChEs. The results give an insight of the mechanism that some OP insecticides can selectively kills Es while without distinct effect on its host, B. mori.

• The Plant Pathology Journal
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• 제37권5호
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• pp.437-445
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• 2021

Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What's more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

• Journal of Microbiology
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• 제45권2호
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• pp.133-138
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• 2007

This study addresses the susceptibility of Spodoptera frugiperda (Sf9 and Sf21), Trichoplusia ni (Hi5), and S. exigua (Se301) cells to the Bombyx mori nucleopolyhedrovirus (BmNPV). Although these cells have classically been considered nonpermissive to BmNPV, the cytopathic effect, an increase in viral yield, and viral DNA synthesis by BmNPV were observed in Sf9, Sf21, and Hi5 cells, but not in Se301 cells. Very late gene expression by BmNPV in these cell lines was also detected via ${\beta}-galactosidase$ expression under the control of the polyhedrin promoter. Sf9 cells were most susceptible to BmNPV in all respects, followed by Sf21 and Hi5 cells in decreasing order, while the Se301 cells evidenced no distinct viral replication. This particular difference in viral susceptibility in each of the cell lines can be utilized for our understanding of the mechanisms underlying the host specificity of NPVs.

• Food Science and Biotechnology
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• 제17권2호
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• pp.219-227
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• 2008

Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.200-203
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• 1994

Spodoptera frugiperda IPLB-SF-21-AE cells were cultivated in a spin-filter bioreactor with continuous perfusion for the recombinant $\beta$-galactosidase production. At the perfusion rate of 0.06 $hr^{-1}$, the maximum cell density of insect cells in this bioreactor system reached 3.5$\times$$l0^6$ viable cells/ml using the Grace media containing 5% FBS and 0.3% Pluronic F-68. The recombinant $\beta$-galactosidase production of 8, 100 units per reactor volume was also achieved at this perfusion rate.

• Archives of Pharmacal Research
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• 제23권5호
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• pp.477-481
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• 2000

The cytotoxicity of crude insect drugs was measured using HeLa cells originating from human cervix and uterine cancer. using the dye uptake assay in order to find potential anticancer agents. Three kinds of extracts (buffer, methanol and ethylacetate) were prepared from 26 insects and used as raw materials for the activity assay. Among these, the buffer extracts from Tabanus, Mylabris and Huechys showed a potent anticancer activity, and those from Catharsius, Red ant, Scorpion, Tabanus and Vespae Nidus showed a strong L-amino acid oxidase (AAO) activity as well as cytotoxicity. In contrast, buffer extracts from Gryllotalpa orientalis and Apriona germari larvae showed greater/more rapid Hela cell growth than that of other insects.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
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• pp.70-70
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• 1996

The cytotoxicological effect of Bt 1-endotoxin, well-known as a bioinsecticide, was investigated on ion channel of insect cell lines. This study attempted to evaluted the specificity by simple experiment to measure the cell swelling using lepidopteran cell lines in isotonic solution containing only one cation. Cell swelling was stimulated in KCI-sucrose isotonic solution as well as TC-100 media containg in solubilized crystal 5-endotoxin. It suggested that the cell swelling by Bt toxin have a relation to K+ channel. The cell swelling may be due to the stimulation K+ influx and simultaneously the penetration of H2O induced by Bt toxin, because the stimulation of swelling was observed with the solubilized toxin in KCI-sucrose isotonic solution, but not in sucrose isotonic solution. Moreover the specific K+ channel blocker, such as 4-arnjnopyrimidine(4-AP) and ouabain, showed the significant effect on the cell swelling induced by Bt toxin. The increasement of the cell swelling induced by 4-AP suggested to be caused by the block of K+ efflux through K+ leak channels. The inhibition of cell swelling by ouabain, which is the well-known inhibitor of Na+, K+-ATPase, suggested to be due to decreasement of K+ influx following diminishment of Na+, K+-ATPase activities.

• 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.