• 한국미생물·생명공학회지
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• 제21권5호
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• pp.504-510
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• 1993

The effect of AcNPV infection conditions such as serum concentration, pH, CaCl2, lysosomotropic agent, cell density at infection, agitation, aeration and nutritional supplementattion on recombinant protein production in Spodoptera frugiperda 21 cells was investigated using tissue culture flask, bottle and spinner flask. It was shown that serum, CaCl2, pH and cell density at infection affected recombinant production. The lysosomotropic agent did not significantly influence recombinant protein production.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.361-364
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• 1999

The binding characteristics of Anagrapha falcifera nuclear polyhedrosis virus (AtNPV) to Spodoptera frugiperda 21 (Sf21) cells were investigated. The cells displayed an affinity of 4.7×10/sup 10/M/sup -1/ with about 3,300 binding sites per cell. The biochemical nature of the AfNPV-binding sites on the cell surface was also partially identified. Our findings suggest that the binding-site moiety has a glycoprotein component, but that the direct involvement of oligosacccharides containing N-acetylglucosamine or sialic acid residues in binding is unlikely, and that AfNPV entry into Sf21 cells may be via receptor-mediated endocytosis.

• 대한의생명과학회지
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• 제11권1호
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• pp.57-61
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• 2005

The number of sites at which a protein can be readily cleaved by a proteolytic enzyme is greatly influenced by its three-dimensional structure. For native, properly-folded proteins both the rate of cleavage and number of sites at which cleavage takes place are usually much less than for the denatured protein. In order to compare the tertiary structure of recombinant HepG2 type glucose transporter with that of its native counterpart in the erythrocyte, the pattern of tryptic cleavage of the protein expressed in insect cell membranes was therefore examined. After 30 minutes digestion, a fragment of approximate Mr 19,000-21,000 was generated. In addition to this, there were two less intensely stained fragments of apparent Mr 28,000 and 17,000. The pattern of labelling was similar up to 2 hours of digestion. However, the fragments of Mr 19,000-21,000 and Mr 17,000 were no longer detectable after 4 hours digestion. The observation of a very similar pattern of fragments yielded by tryptic digestion of the HepG2 type transporter expressed in insect cells suggests that the recombinant protein exhibits a tertiary structure similar if not identical to that of its human counterpart. Also, the endogenous sugar transporter(s) present in Sf21 cells did not bind cytochalasin B, the potent transporter inhibitor. Therefore, the baculovirus/Spodoptera frugiperda (Sf) cell expression system could be very useful for production of large amounts of human glucose transporters, heterologously.

• 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.

• 환경생물
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• 제40권3호
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• pp.281-289
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• 2022

본 연구에서는 온도에 따른 열대거세미나방의 산란 특성을 조사하였다. 18, 21, 24, 27, 30℃에서 열대거세미나방의 암컷 성충의 수명기간은 각각 19.2일, 22.4일, 20.4일, 19.0일 및 13.9일이었으며, 산란 전 기간은 각각 5.2일, 4.9일, 5.2일, 5.3일 및 3.5일이었고, 총산란 수는 각각 887.4개, 1,246.4개, 1,348.9개, 1,154.9개 및 1,034.2개였다. 온도별 열대거세미나방의 암컷 생존율은 18℃에서 13일 이후, 21℃에서 14일 이후, 27℃에서 15일 이후 및 24℃와 30℃에서는 9일 이후 급속하게 감소하였다. 산란 시작 후 3일째에 전체 산란의 50%가 이루어졌고, 18~24℃에서는 산란시작 후 7일째, 27~30℃에서는 5일째에 전체 산란의 90%가 완료되었다. 10엽기 이하의 옥수수 포장에서 열대거세미나방 성충은 하위엽에 41.4%, 중위엽에 46.8% 및 상위엽에 11.7%를 산란하였으며, 산란된 난괴의 66.7%가 잎 뒷면에 산란되었다. 열대거세미나방 성충을 5월 12일, 5월 17일, 5월 25일 및 5월 30일에 방사한 결과, 난괴당 알 수는 각각 89.9개, 88.5개, 126.6개 및 127.9개였다. 접종 후 세대성충의 산란은 6월 하순부터 관찰되었으며, 6월 하순에 산란된 난괴당 알 수는 155.8개, 7월 상순에 270.7개 및 7월 중순에 303.5개였다. 본 연구에서 보고한 열대거세미나방의 산란 특성은 성충 비래 시기에 따른 발생 예측 분석 및 방제 대책을 마련하는 데 활용될 수 있을 것이다.

• 한국응용곤충학회지
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• 제61권2호
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• pp.349-356
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• 2022

열대 및 아열대성 비래해충으로 최근 국내에도 옥수수에 피해를 주고 있는 열대거세미나방의 온도별 발육특성을 인공먹이를 이용하여 사육하며 조사하였다. 18, 21, 24, 27, 30, 32℃ 항온조건에서 알에서 성충까지의 발육하기까지 각각 79.8, 54.2, 34.3, 28.4, 24.6, 24.0일이 소요되어 온도가 증가할수록 발육기간이 짧아졌다. 암컷 번데기의 발육기간은 수컷보다 짧았다. 유충은 보통 6령까지 발육하였으나 저온에서 7령 이상의 비율이 증가하였다. 온도에 따른 발육은 직선회귀에 부합하였으며, 직선회귀식을 이용하여 각 발육태별 발육영점온도와 유효적산온도를 분석한 결과, 알은 12.9℃와 37.0 DD, 유충은 11.3℃와 286.3 DD, 번데기는 12.6℃와 132.2 DD, 알에서 성충까지는 11.8℃와 456.8 DD 였다.

• 대한의생명과학회지
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• 제12권1호
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• pp.17-22
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• 2006

The baculovirus expression system is a powerful method for producing large amounts of the human erythrocyte-type glucose transport protein, heterologously. Characterization of the expressed protein is expected to show its ability to transport sugars directly. To achieve this, it is a prerequisite to know the properties of the endogenous sugar transport system in Spodoptera frugiperda Clone 21 (Sf21) cells, which are commonly employed as a host permissive cell line to support the baculovirus replication. The Sf21 cells can grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transport system. However, unlike the human glucose transport protein that has a broad substrate and inhibitor specificity, very little is known about the nature of the endogenous sugar transport system in Sf21 cells. In order to characterize further the inhibitor recognition properties of the Sf21 cell transporter, the ability of phloretin, cytochalasin B and D-fructose to inhibit 2-deoxy-D-glucose (2dGlc) transport was examined by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. The 2dGlc transport in the Sf21 cells was very potently inhibited by phloretin, the aglucone of phlorizin with a $K_i$ similar to the value of about $2{\mu}M$ reported for inhibition of glucose transport in human erythrocytes. However, the Sf21 cell transport system was found to differ from the human transport protein in being much less sensitive to inhibition by cytochalasin B (apparent $K_i$ approximately $10\;{\mu}M$). In contrast, It is reported that the inhibitor binds the human erythrocyte counterpart with a $K_d$ of approximately $0.12\;{\mu}M$. Interestingly, the Sf21 glucose transport system also appeared to have high affinity for D-fructose with a $K_i$ of approximately 5mM, contrasting the reported $K_m$ of the human erythrocyte transport protein for the ketose of 1.5M.

• 한국농림기상학회지
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• 제25권3호
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• pp.142-150
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• 2023

기후온난화가 지속되면서 국내에도 열대거세미나방의 침입 시기가 빨라지고 유입량도 증가하여 작물에 대한 피해도 크게 증가하고 있는 실정이다. 본 연구는 열대거세미나방에 의한 작물의 피해를 최소화하고자 옥수수 포장에 요방제 수준을 통한 방제시기를 설정하고 사료용 옥수수 포장에서 약제 살포 방법에 따른 방제효과를 조사하였다. 옥수수 출사기에 피해율 4% 조건에서도 피해과율은 70%로 높은 피해 양상을 나타내었다. 열대거세미나방 2령 유충의 경제적 피해수준은 주당 0.7마리로 나타났으며, 요방제 수준은 0.6마리로 나타났다. 옥수수 도매 단가를 적용하여 소득을 산출한 결과, 피해율 4% 조건에서도 895,221원/10a 손실액이 났으며, 피해율이 높아질수록 소득 감소는 매우 큰 것으로 나타났다. 이를 방제하기 위하여 열대거세미나방에 등록된 단제 10종에 대해 살충효과를 검정한 결과, 4종 약제(에마멕틴벤조에이트, 클로란트라닐리프롤, 인독사카브, 스피네토람)에 대해 93.3% 이상의 높은 살충 활성을 나타내었으며, 약효 지속성 검정을 통해 3종 약제(클로란트라닐리프롤, 인독사카브, 스피네토람)에서 높은 잔류 효과로 열대거세미나방의 방제에 효과적인 것으로 생각된다. 따라서 열대거세미나방에 높은 활성을 나타내는 인독사카브 액상수화제, 클로란트라닐리프롤 입상수화제를 대상으로 사료용 옥수수의 출사 전에 관행방제와 항공방제를 각각 7일 간격으로 2회 실시하였다. 그 결과, 1차 방제를 통해 항공방제보다 관행방제에서 인독사카브 액상수화제 46.3%p, 클로란트라닐리프롤 입상수화제 21.7%p의 높은 방제가를 나타내었다. 2차 방제에서도 항공방제보다 관행방제에서 인독사카브 액상수화제 26.7%p, 클로란트라닐리프롤 입상수화제 40.4%p의 높은 방제가가 조사되었다. 이에 따라 인력방제 대체 수단을 많이 사용하는 최근 상황에서 방제 효과를 제고할 수 있는 방안 마련이 필요한 것으로 사료된다.

• 대한의생명과학회지
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• 제11권4호
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• pp.487-492
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• 2005

The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.

• 대한의생명과학회지
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• 제9권4호
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• pp.177-181
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• 2003

The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.