• Microbiology and Biotechnology Letters
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• v.21 no.5
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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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• v.9 no.3
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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.

• Biomedical Science Letters
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• v.11 no.1
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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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• v.4 no.3
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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.

• Korean Journal of Environmental Biology
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• v.40 no.3
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• pp.281-289
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• 2022

The effect of five different constant temperatures (18, 21, 24, 27, and 30±1℃) and a photoperiod of 14 : 10 (L :D) h on the reproduction parameters of Spodoptera frugiperda was studied. The longevity of adult female S. frugiperda decreased with increasing temperature (22.4 days at 21℃ and 13.9 days at 30℃) but not at 18℃. The pre-oviposition period and oviposition period was the shortest at 30℃ compared to the other temperatures. The total fecundity egg count was 887.4, 1,246.4, 1,348.9, 1,154.9, and 1,034.2 at 18, 21, 24, 27 and 30℃, respectively, during its life span. The survival rate of female S. frugiperda decreased rapidly after 13 days at 18℃, after 14 days at 21℃, after 15 days at 27℃, and after 9 days at 24℃, and 30℃. On the third day after the start of oviposition, 50% of the total fecundity was accomplished. In corn fields at less than the 10-leaf stage, the distribution of S. frugiperda egg masses was observed in the middle and lower plant regions, corresponding to 46.8% and 41.4% of the total egg masses, respectively. Egg masses were mostly found on the underside of the leaf blade (abaxial) of corn(66.7%). After releasing S. frugiperda adults on May 12, May 17, May 25, and May 30, the number of eggs per egg mass was 89.9, 88.5, 126.6, and 127.9, respectively. Egg masses of the subsequent generations of S. frugiperda were observed from late June, and the number of eggs per egg mass was 155.8 in late June, 270.7 in early July, and 303.5 in mid-July.

• Korean journal of applied entomology
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• v.61 no.2
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• pp.349-356
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• 2022

Fall armyworm, Spodoptera frugiperda, is a invasive migratory pest of native to the tropical and sub-tropical regions that has recently invaded Korea with damage to cornfield. The study was conducted to investigate the development periods of S. frugiperda on artificial diet at six different temperatures. The developmental period from eggs to adult at 18, 21, 24, 27, 30 and 32℃ was 79.8, 54.2, 34.3, 28.4, 24.6 and 24.0 days, respectivery and decreased with increasing temperature. The pupal periods of females were shorter than males. Most of the larvae developed through six instar, but the ratio of 7 or more instar increased at low temperatures. The relationship between the development rate and temperature was fitted with by linear regression analysis. The lower development threshold for egg and larva development was 12.9℃ and 11.3℃, respectively, 12.6℃ for pupae and 11.8℃ for egg-to-adult development. The effective accumulative temperature for the development of the respective life cycle stages were 37.0 DD for eggs, 286.3 DD for larvae, 132.2 DD for pupae and 456.8 DD egg-to-adult development.

• Biomedical Science Letters
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• v.12 no.1
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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.

• Korean Journal of Agricultural and Forest Meteorology
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• v.25 no.3
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• pp.142-150
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• 2023

As global warming continues, the time of invasion of Spodoptera frugiperda has been advanced and the inflow rate has been increasing, leading to great increases in damage to crops. In this study, in order to minimize crop damage caused by S. frugiperda, the control period was set for corn fields through control thresholds, and the control effects according to the chemical spraying methods were investigated in forage corn filed. Even under the condition of 4% injury level during the corn silking stage, the damage rate of ear was 70%, showing an aspect of extensive damage. The economic injury level of S. frugiperda second instar larvae was shown to be 0.7 larvae per stalk, and the control threshold level was shown to be 0.6 larvae. The income was calculated by applying the corn wholesale unit price, and according to the result, even under the condition of injury level of 4%, there was a loss of KRW 895,221/10a, and the higher the injury level, the greater the decrease in income. To control S. frugiperda, the insecticidal effects of 10 single formulations registered for S. frugiperda were tested, and according to the results, four types(emamectin benzoate, chlorantraniliprole, indoxacarb, and spinetoram) showed high insecticidal activity not lower than 93.3%, and three types (chloran- traniliprole, spinetoram, and indoxacarb) were considered to be effective in controlling S. frugiperda as they showed high residual effects through insecticidal effect persistence tests. Therefore, conventional control and aerial control were conducted twice at 7-day intervals with indoxacarb SC and chlorantraniliprol WP, which show high activity against S. frugiperda, respectively, prior to the silking of forage corn. As a result, conventional control showed higher control values, 46.3%p in the case of indoxacarb SC and 21.7%p in the case of chlorantraniliprol WP, than aerial control through the primary control. In the secondary control too, higher control values of 26.7%p in the case of indoxacarb SC and 40.4%p in the case of chlorantraniliprol WP were found in conventional control than in aerial control. Therefore, it is considered necessary to prepare measures to improve the control effects in the recent situation where alternative methods for manpower control are widely used.

• Biomedical Science Letters
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• v.11 no.4
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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.

• Biomedical Science Letters
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• v.9 no.4
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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.