• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.145-149
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• 2003

ApolipophorinIII (apoLp-III) is a protypical exchangeable apolipoprotein that is abundant in hemolymph of many insect species. Its function lies in the stabilization of low-density lipophorin particles (LDLp) crossing the hemocoel in phases of high energy consumption to deliver lipids from the fat body to the flight muscle cells. But, recent studies with naive Galleria mellonella-apoLp-III gave first indication of an unexpected role of that protein in insect immune activation. In this research, we cloned a cDNA encoding putative apoLp-III from the silkworm, Bombyx mori injected with E. coli and characterized its role. We constructed a cDNA library using whole bodies of B. mori larvae injected with E. coli, carried out the differential screening, and selected the up-regulated clones. Among these clones, we focused on a cDNA showing a high sequence similarity to the apolipophorinIII from other insects and analyzed the nucleotide and deduced amino acid sequences. The pupative B. mori Jam123 apoLp-III cDNA contained 1,131 bp encoding 186 amino acid residues. Phylogenetic analysis revealed that the nucleotide and amino acid sequences of the B. mori apoLp-III cDNA formed a highly inclusive subgroup with Bombycidae. But, it was interesting that B. mori Jam123 is closer to B. mandarina than B. mori P50 and B. mori N4. Northern blot analysis showed a signal in the fat body, posterior silkgland and midgut.

• Current Research on Agriculture and Life Sciences
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• v.20
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• pp.55-63
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• 2002

This experiment was to know properties of Sericin Jam that development, growth of silkgland, content of sericin and composition of amino acid in hemolymph. The characters of Sericin Jam can he seen form this experiment. Hatching ratio was 85% in Sericin Jam and 95% in Jam 120. Especially hatching period of Sericin Jam was longer than Jam 120 and also hatching of Sericin Jam was ununiform. The larval duration of Sericin Jam was 20 days and 23 hours, and Jaw 120 was 21 days and 22 hours. In Sericin Jam, middle silkgland contain a great p arts in silkgalod and posterior silkgand is short and no curves. The period of mounting to emergence was 12 days in Sericin Jam. The period of pupa was 7day. It is property of Sericin Jam that the period is short. Cocoon was very thin and light in Sericin Jam. Weight of cocoon shell of Sericin Jam is 2.7cg. The sericin protein quantity was 100% in cocoon shell of Sericin Jam, about 28% in cocoon shell of Jam 120, however the sericin ratio per Sericin Jam cocoon was 34.6% compare to cocoon shell of Jaw 120 in sericin protein quantity. The amino acids in hemolymph of Sericin Jaw was much hidtidine, lysine, glut amic acid. And the amount of almost amino acids were increased depends upon development at t he 5th instar.

• Journal of Sericultural and Entomological Science
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• v.23 no.2
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• pp.37-49
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• 1982

Larvae of the silkworm (Bombyx mori L.) were reared during the 5th instar on the four kinds of artificial diets on the basis of the different amounts of soybean meal used as the protein source. In this experiment it was shown that the various levels of protein in the diet affected not only the growth and silk production but the digestibility of the diet. haemolymph protein and uric acid excretion. The results obtained are summarized as follows; 1. By an increase of the level of protein in the diet the apparant digestibility was increased. but the protein digestibility was comparatively decreased. 2. Larval body weight increment was not observed by the 3rd day of the 5th instar, but was increased from the 4th day as the level of protein was increased in the diet. 3. After the 3rd day of the 5th instar, protein content in the hemolymph was increased steeply by an increase of the protein content in the diet. However, the percentage of hemolymph protein to the ingested protein was decreased from the 2nd day of the 5th instar and increased more or less after the 4th day. 4. An increase of the uric acid excretion was observed as the content of protein in the diet was increased but the pattern of the uric acid excretion was different between high and low-protein diet. However, the percentage of the uric acid excretion to the ingested protein and to the hemolymph protein were both decreased steeply after the 2nd day of the 5th instar. 5. It was also evident that the high-protein diet increased the cocoon productivity. 6. It showed that the feed efficiency for body weight increment and silk formation was high by an increase of the level of protein in the diet, but the protein efficiency was not.

• Journal of Sericultural and Entomological Science
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• v.43 no.2
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• pp.59-66
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• 2001

This study was conducted to elucidate phylogenetic relationships and genetic characterization of silkworms that might be recognized as the Korean native strains. Genetic characterization in isozymes and the proteins of larval hemolymph of 17 silkworms were observed by acrylamide gel eletrophoresis, on 12 genes; Bph, Bes, les, Amy-hc, Ict-A, -B, -D,-E,-H, Pfl, Pst, Lp. Gene frequencies in each locus were compared other geographic strains. Korean native strains were remarkably different from others considered as the genetic characterization of Korean native strains. Phylogenetic relationships in Korean native strains were analysed using RAPD-PCR markers. A total of 40 primers were used and 346 bands of amplified DNA were generated from geographic strains. Genetic similarity based on the RAPD bands was used to construct phylogenetic dendrogram based on analysis of bard sharing data of amplified markers. Genetic similarity ranged from 0.595 to 0.860. In the genetic relationship based on dendrogram, they were classified into Bombyx mori group (including 16 domesticated silkworm strains) and B. mandarina group. The Bombyx mori group was separated into three sub-groups at the genetic similarity of 0.6930, including Korean, Japanese and Chinese groups. According to this result, the Korean native variety can be considered as a clearly different variety from other geographic strains. It may be concluded that the Korean native strains are also one of original geographic variety such as Japanese, Chinese, etc.

• KSBB Journal
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• v.11 no.4
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• pp.438-444
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• 1996

Using recombinant Vaccinia virus(vSC8) that express ${\beta}$-galactosidase, a model heterologous protein, conditions for virus and protein production were investigated in tissue culture flask. As host animal cells HeLa and HeLa S3 were used. It was demonstrated that cells infected during the exponential growth phase gave higher protein yield than those infected during the stationary growth phase and calf serum concentration after virus infection did not significantly alter protein yield. Pretreatment of cell layer with hypotonic solution enhanced the virus infectivity. Optimum cell growth and recombinant protein production was achieved at $37^{\circ}C$. But, during 2 hours of virus infection period incubation temperature must be lowered to 20∼$30^{\circ}C$ for maximum recombinant protein yield. To enhance virus replication, the effects of adrenal glucocorticoid hormone (Dexamethasone) and silkworm hemolymph were evaluated. Only dexamethasone increased about 20% of ${\beta}$-galactosidase yield in HeLa S3 cells when added with 10-7∼10-5M concentration 24 hours before infection.

• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.225-232
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• 1998

We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

• 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.