• Journal of Sericultural and Entomological Science
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• v.31 no.2
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• pp.72-81
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• 1989

Antheraea yamamai vitellin was purified from matured eggs by polyacryamide gel electrophoresis, also stage dependent appearance, immunological comparison and relative content of the protein were investigated. 1. Vitellogenin, the precursor of vitellin, was first detected in the larval hemolymph at the late spinning stage by polyacrylamide gel electrophoresis and immunoelectrophoresis. 2. The electrophoretic mobility of the vitellin was identical with that of Bombyx mori and of Bombyx mandarina. However, the specific antiserum against A. uamamai vitellin did not react with either that of Bombyx mori or Bombyx mandarina in immumo-diffusion test. 3. Relative content of A. yamamai vitellin to the total soluble egg protein was 46.0 percent and did not change till eight days after oviposition. But the content started to decline from ten days after oviposition and was negligible in the five or serventeen month old eggs.

• Journal of Sericultural and Entomological Science
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• v.31 no.2
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• pp.82-90
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• 1989

Antheraea yamamai vitellin was purified from matured eggs by polyacrylamide gel electrophoresis for characterization of its biochemical properties : molecular weight, sugar and lipid composition, amino acid composition and electron microscopic morphology, etc. 1. A yamamai vitellin was composed of two subunits, large and small, showing different mobility in SDS-polyacrylamide gel electrophoresis. 2. The molecular weight of the vitellin was estimated to be approximately 450,000 dalton and the large and small subunits were 174,000 dalton and 44,000 dalton, respectively. 3. The vitellin seemed to be a glycolipoprotein since it showed a positive reaction to coomassie brilliant blue, sudan black B and PAS staining. Both subunits were similiar in this aspect. 4. Lipid of the witellin reveraled several different types including saturated lipids. 5. When the vitellin was incubated at 7$0^{\circ}C$ for 60 minites its apoprotein still cross-reacted to the specific antiserum to the native vitellin. Its sugar components were also detected by PAS staining, but its lipid portion was not detected by sudan black B staining. 6. Its amino acid composition was similar to that of other insects, but its glycine content was peculiarly very high. 7. The vitellin molecule was spherical in shape with a diameter of 14$\pm$0.8nm by negatively.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.1
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• pp.55-59
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• 2001

The vitellin of the red flour beetled Tribolium castaneum Herbst was purified and characterized. The vitellin of T. castaneum was purified by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In native-polyacrylamide gel electrophoresis, vitellin of T. castaneum was detected as a single band. This native vitellin has molecular weight of 440 kDa. The vitellin of T. castaneum is composed of three polypeptides, designated Vnl (178 kDa), Vn2 (168 kDa) and Vn3 (52 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph and egg extracts, but not observed in the male. These three polypeptides gradually decreased during embryogenesis. Polyclonal antiserum raised against purified vitellin reacted with the three polypeptides, Vnl, Vn2 and Vn3. Antisera raised against Vn1 and Vn2 cross-reacted with the two large subunits, Vnl and Vn2, respectively. Another subunits Vn3, however, was not cross-reacted with these two antisera. Also, antiserum raised against Vn3 did not cross-react with the Vn1 and Vn2.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.131-135
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• 2000

The vitellin of the fireflies, Luciola unmunsana and L. lateralis was characterized. The vitellin of L. unmunfon is composed of two subunits, designated Vnl (195 kDa) and Vn2 (185 kDa) in SDS-polyacryamide gel electrophoresis. These two subunits of vitellin of L. unmunsana gradually decreased during embryogenesis. As expected, these protein bands were presented in female adult hemolymph and egg extracts, but not in male. The vitellin of L. lateralis is also composed of two subunits, designated Vnl (195 kDa) and Vn2 (180 kDa) in SDS-PAGEi and these two protein bands gradually decreased during embryogenesis. Western blot analysis using each of polyclonal antiserum against vitellins of L. unmunsana and L. lateralis showed that two antisera strongly crossereacted with vitellin subunits of L. unmunsana and L. lateralis, suggesting that vitellins of L. unmunsana and L. lateralis have similarity with each other.

• Journal of Sericultural and Entomological Science
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• v.30 no.2
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• pp.96-104
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• 1988

Vitellin, the major yolk protein of the silkworm, Bombyx mori was pruified, and its immunological properties and the quantitative changes during embryogenesis were studied. The ovary transplantation into male hosts was also carried out to find its effect on the yolk protein synthesis. The pupal vitellogenin and the egg vitellin of Bombyx mori were purified by DEAE-cellulose column chromatography. These two female specific proteins showed the same mobility in the polyacrylamide gel electrophoresis and the same reaction in the double immunodiffusion test. The immunological identity was also observed between the vitellins of Bombyx mori and Bombyx mandarina. The rudimentary ovaries transplanted into the male hosts of silkworms produced eggs without vitellin, indicating that the yolk precursors synthesized in other female organ beyond the ovary were necessary to produce vitellins. The major yolk protein, vitellin was disintegrated and utilized mostly during late stage of embryogenesis. It was different characteristics from the egg specific protein, which was utilized continuously from the early embryonic stage.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.167-172
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• 2001

The vitellin of firefly, Pyrocoelia rufa, is composed of three polypeptides, designated Vn1 (175 kDa), Vn2 (160 kDa) and Vn3 (45 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph, ovary and egg extracts, but not observed in the male. This vitellin was purified from the eggs of P. rufa by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In nature, vitellin of P. rufa has molecular weight of 400 kDa. Western blot analysis using polyclonal antiserum against purified vitellin showed that the antiserum was reacted with the three polypeptides, Vnl, Vn2 and Vn3 from the female adult hemolymph, ovary and egg extracts. Amino acid residues at N-terminus of three subunits were sequenced. The N-terminal sequences of large subunits, Vnl and Vn2, were similar to each other, But, the N-terminal sequences of small subunits Vn3, did not have any signnificant homology with large subunits.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.473-479
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• 1997

The yolk protein of spotted flounder, Verasper variegatus was purified by precipitation wit cold distilled water, followed by Sepharose CL-6B column chromatography. The purified protein was identified as vitellin by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The purified vitellin from ovarian crude extracts has same antigenic determinants with the female specific serum protein, vitellogenin. The molecular weight of purified vitellin was estimated about 550 kD by gel filfration. The vitellin was composed of three major subunits with molecular weight of about 108, 85 and 31 kD, and two minor subunits. The vitellin was identified by western blot analysis using anti-vitellin antibody.

• Journal of Sericultural and Entomological Science
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• v.29 no.1
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• pp.20-30
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• 1987

In the hemolymph proteins of the silkworm reared with mulberry leaves and artificial diet, electrophoretic patterns of Storege Protein 1 (SP1) and Vitellogenin(VG), changes of relative concentration of SP1, JHA effects during the developmental stages, and estimation of the molecular weight of vitellin subunits were investigated in the present study. 1. Storage Protein 1 (SP1) showed female specificity from the middle stage of the fifth instar to the end of spinning stage in the silkworm reared with mulberry leaves and artificial diet. 2. Vitellogenin (VG) showed female specififcity just afrter pupation in the silkworm reared with mulberry leaves and artificial diet. 3. Storage Protein 2 (SP2) without female specificity was detected from the early stage of the fifth instar to the first or second day after pupation. 4. Vitellin was composed of two non-identical subunits (vitellin-heavy chain ; VTL-H, vitellinlight chain ; VTL-L) with mplecular weight of 186,000 and 41,000. Also, there were no differenes between the molecular weights of vitellin subunits obtained from silkworms reared with mulberry leaves and artificial diet. 5. Relative concentration of Storage Protein 1 (SP1) after topical application - 0.5, and 10$\mu\textrm{g}$ per body weight - JHA on the 60th hour of the fifth instar showed the highest increase with the treatment of 5$\mu\textrm{g}$ and a higher increase with the treatment of 10$\mu\textrm{g}$ compared with the control (no topical application of JHA).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.432-442
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• 1992

To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

• Journal of Sericultural and Entomological Science
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• v.33 no.1
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• pp.1-5
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• 1991

PBS soluble- and insoluble-extracts form silkworm eggs were analysed by native-PAGE and SDS-PAGE. During the embroyonic development, soluble proteins which are mainly composed of ESP, vitellin and 30K proteins showed similar degradation pattern in both electrophoretic analyses. Several peptides which seemed to be intermediated forms of yolk proteins were detected in latter part of embryogenesis, while a protein band newly appeared in one day elapsed after acid treatment. When insoluble extracts from silkworm eggs were analysed with SDS-PAGE, several peptides were detected at the later stages of the embryonic development, and newly hatched larvae. These peptides are considered to be structure proteins for embryonic morphogenesis.