• Journal of Sericultural and Entomological Science
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• v.49 no.2
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• pp.47-50
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• 2007

Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.

• Journal of Sericultural and Entomological Science
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• v.37 no.2
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• pp.181-185
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• 1995

To enhance expression of foreign gene by the novel expression vector, pBmKSK1, of Bombyx mori nuclear polyhedrosis virus, E. coli $\beta$-galactosidase gene expressing recombinant virus was infected in BmN-4 cells and various concentrations of silkworm hemolymph were added to the recombinant virus-infected BmN-4 cells containing fetal bovine serum. The expression efficiency of foreign gene was determined by $\beta$-galactosidase activity in the culture media. The results showed that the silkworm hemolymph was effective to expression of foreign gene in the BmN-4 cells, suggesting that the silkworm hemolymph could be substituted for fetal bovine serum in the BmN-4 cells to enhance expression of foreign gene.

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.119-123
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• 1994

By in vivo labelling of AMHP using[35S]-methionine, fat body culture and immunological analysis, it is proved that Bombyx adult fat body synthesizes 18K and 20K subunits of AMHP and releases them into haemolymph. Also these peptides are assembled to form native AMHP in the adult haemolymph.

• Parasites, Hosts and Diseases
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• v.28 no.1
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• pp.11-24
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• 1990

In order to observe the antigenic localization in the tissues of the adult Clonorchis sinensis, immunogold labeling method was applied using serum immunoglobulins (IgG) of either worm·infected rabbits (group I) or antigen-immunized rabbits (group II) (by the body quid obtained from the adult worms). The electron micrographs of the sectioned worm tissue antigens, embedded in Lowicryl HM 20 medium and stained with protein A-gold complex (particle sixte: 12 nm), were compared between the group I and group II. The gold particles were observed in the interstitial matrix of the worm parenchyma, the epithelial lamellae of the cecum, and the cecal lumen both in group I and II. But the particles were in general more concentrated in group II. The gold particles were not observed on the basal lamina of the tegument or on vitelline glands in group I, while they were highly concentrated on those areas in group II. There were also differences in the antigenicity of interstitial matrix(reacted with group I IgG) and head part(reacted with group II IgG) of the sperm cells in the seminal receptacle. Conclusively, it is suggested that the substances comprising the basal lamina of the tegument or vitelline glands act as specific antigens reacting with antigen(body quid) immunized rabbit IgG. On the other hand, the substances in the cecal lumen and cecal epithelial lamellae are thought to be the specific antigen that react with the worm-infected rabbit IgG.

• Korean journal of applied entomology
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• v.43 no.1
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• pp.21-26
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• 2004

These studies were carried out to investigate the oviposition and host feeding characteristics of Neochrysocharis formosa Westwood, an endoparasitoid of Liriomyza trifolii. N. formosa female oviposited inside the abdomen at the rear end of the host larva. Number of eggs and host feeding per day of N. formosa tended to increase as temperatures go up. The ratio of oviposition and host feeding of N. formosa were 0.5, 0.9, 1.0 and 1.0 at 15, 20, 25, and 30$^{\circ}C$, respectively. In 25$^{\circ}C$, number of eggs and host feeding were 176.0 and 188.7, respectively, and aduly female longevity was 11.8 days. N. formosa female preferred to the 3rd rather than 1st and 2nd larva for oviposition and host feeding.

• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.30-36
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• 1986

Polyacrylamide gel electrophoresis and autoradiography were applied to investigate the developmental profiles of the major hemplymph proteins (MHPs) and their biosynthesis. In addition, some biochemical methods were also used to isolate and purify the MHPs. The obtained results are summarized as follows. 1. MHP-a began to appear from the 2nd day of the fourth-instar larva while MHP-b and -c were detected first on the 1st day of the fifth-instar larva. All these proteins, however, showed a drastic increase in concentration at the 2nd day of the fifth-instar larva. 2. MHP-b and -c were synthesized in fat body on early day of the fifth-instar larva, but the possibility of MHP-a synthesis in fat body was excluded. 3. MHP-b was isolated and purified by heat-treatment (6$0^{\circ}C$), gel filtration on Sephadex G-100 and column chromatography on DEAE-cellulose and CM-cellulose. Purified MHP-b showed a single band on polyacrylamide gel- and SDS-polyacrylamide gel electrophoresis.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2001.04a
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• pp.69-69
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• 2001

• Journal of Sericultural and Entomological Science
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• v.30 no.1
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• pp.25-32
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• 1988

In order to examine a physiological role of juvenile(JH) binding proteins in the hemolymph of the silkworm larva, Bombyx mori, [3H] JH I incubated hemolymph was separated by polyacrylamide gel electrophoresis in the fifth-instar larva and the activity of the binding protein was analyzed using charcoal binding assay. The results obtained were as follows; 1. The JH was bound by two protein fractions in the hemolymph of the fifth-instar larva; One was JH binding lipoprotein(JH-LP), the other was JH speific binding protein(JHBP). Their relative mobility values(Rm) were 0.3∼0.33 and 0.81∼0.84, respectively. There were no valid differences in those values from developmental stages of both male and female silkworms. 2. Total protein contents of the hemolymph were gradually increased during the fifth-instar larva, while at the prepupa decreased. The maximum ones were observed at the spinning period and the contents from female were much higher than those from the male. 3. JH binding activity per ml of the hemolymph was low in the early stage of the fifth-instar larva and its activity was maximized at the psinning period and at the prepupa slightly decreased. 4. There was a similar pattern between changes of the JH binding activity per ml of the hemolymph and of the total protein contents of the hemolymph. 5. The JH binding activity per mg of the hemolymph proteins was high in the early stage of the fifth-instar larva, while from the 6th day of the fifth-instar larva to the prepupa its activity showed the lowest levels.

• Journal of Sericultural and Entomological Science
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• v.49 no.1
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• pp.28-32
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• 2007

This study was carried out to investigate the utilization of hemolymph of silkworm, Bombyx mori, as a substitute for fetal bovine serum(FBS) in the insect cell culture. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate for large scale collection. The hemolymph was collected from 5 th instar larva by centrifugation after cutting of the abdominal legs was more appropriate procedure for large scale collection. The cell growth in the medium supplemented with hemolymph(Baekokjam) collected in large scale was almost same as that in the medium hemolymph supplemented with hemolymph collected in small scale. However, the mutant($wE^b$) hemolymph collected in large scale was still less effective in the cell growth, as compared to the Baekokjam hemolymph collected in large scale. The optimum centrifugation condition for large-scale bleeding was 500 rpm and 15 min.

• Journal of Sericultural and Entomological Science
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• v.30 no.1
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• pp.20-24
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• 1988

Adult specific proteins, referred to as ASP-I(adult specific protein of slow mobility) and ASP-II(adult specific protein of slow mobility) at the pharate adult stage of the silkworm, Bombyx mori, were detected by polyacrylamide gel electrophoresis. The adult specific proteins (ASP-I and ASP-II) were defined as a kind of adult hemolymph proteins without sex specificity, and the was no variation in the respective electrophoretic mobility of ASP-I and of ASP-II among the forty-one silkworm varieties. tested.