• The Korean Journal of Zoology
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• v.23 no.1
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• pp.1-12
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• 1980

Changes and possible origin of haemolymph proteins during the cuticle formation and hardening are determined by means of acrylamide gel electrophoresis and immunodiffusion. The results by acrylamide gel electrophoresis showed at least 19 protein bands in the haemolymph and 13 fractions in the fat body with relatively constant pattern during the period of cuticle formation and hardening. Both haemolymph and fat body proteins are generally characterized by the presence of three to four heavy stained bands and several thin bands near the top region of the gel. At least over five haemolymph proteins are constantly present during this period. Immunodiffusion tests show that of total eight to nine pupal haemolymph proteins two proteins were already detected in the fat body before pupation and other two proteins were also found in the fat body immediately after pupation, suggesting fat body as possible source of these two haemolymph proteins.

• International Journal of Industrial Entomology and Biomaterials
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• v.14 no.1
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• pp.37-44
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• 2007

A study was made of the haemolymph protein spectrum of mulberry silkworm (Bomhyx mori L.) from the first larval instar to imago. Horizontal starch gel electrophoresis was used. Sixteen races and eight F1 interracial hybrids, raised in Bulgaria, were analyzed. During the ontogenesis, a total of 17 protein bands (15 cathodic and 2 anodic) were detected. Distinct dynamics in the haemolymph protein spectrum was observed, in result of different expression during the individual development associated with the processes of growth, histolysis and histogenesis. Based on the ontogenetic dynamics found, a correspondence was assumed between some proteins detected by us using the starch gel electrophoresis and major haemolymph proteins (SP1, SP2, MHPs and Vg) detected by other authors using the polyacrilamide gel electrophoresis. Intraracial and interracial polymorphism was observed in four protein zones. The effect of four polymorphic loci with codominant and null alleles was suggested.

• The Korean Journal of Zoology
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• v.26 no.2
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• pp.83-93
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• 1983

Vitellogenin was purified from the haemolymph of female pupae during egg formation of Bombyx mori using DEAE-cellulose column and its quantitative change in various organs with age ws examined by electrophoresis and immunodiffusion. Vitellogenin is distributed in the haemolymph, fat body, and over the egg maturation and especially maintainsa constant level in the haemolymph until just before emergence, indicating that vitellogenin in released into the haemolymph at the same amount as it is taken up by oocytes during pupal period. Immunologically vitellogenin was confirmed to be in the fat body until 7 days after pupation and to undergo a drastic decline thereafter. Also, the interaction of anti-ovary proteins with haemolymph proteins showed at least 3 homogeneous proteins, indicating that other proteins as well as vitellogenin are involved in egg formation.

• Journal of the Korean Society of Tobacco Science
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• v.16 no.1
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• pp.90-96
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• 1994

Changes in haemolymph proteins, hydrolases such as esterase(EST), acid phosphatase(ACP) and alkaline phosphatase(ALP) , and inorganic ion(Na+, K+ and Cl- ) contents were induced by the injection of Bacillus thuringiengis into haemocoel of the last instar larva of Heliothis assulta. Protein concentration of haemolymph was increased until 24 hrs after injection, and decreased thereafter. Among the 8 basic protein bands identified through acid - polyacrylamide gel electrophoresis(PAGE), 2 bands(bands a and b) became stronger by the bacterial infection. Activities of EST and ALP increased until 12 hrs after injection and then fell down, whereas ACP activity was decreased continuously with time after injection. Contents of inorganic ions were all increased by the bacterial injection, showing slow rate of increase in the chloride ion, but rapid in the sodium and potassium ions.

• Journal of Sericultural and Entomological Science
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• v.41 no.2
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• pp.75-81
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• 1999

The storage protein-like protein has been purified from the 5th instar larval haemolymph of the Chinese osk silkwom, Antheraea pernyi, and the preparation was shown to be homogeneous by 7.5% native-PAGE. The molecule was consisted of a single subunit with a molecular weight of 80K, but the number of the subunits was not determined. The protein was defied as glycoprotein by Schiff's regent stining. Rabbit antibody prepared against the purified protein crotein crossreacted with the 5th instar larval haemolymph proteins of Antheraea pernyi and antheraea yamamai, but not with those of Bombyx mori and Bombyx mandarina.

• The Korean Journal of Zoology
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• v.38 no.3
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• pp.305-312
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• 1995

The Injection of viable Escherichia coli K-12 with fifth instar larvae of cabbage butterfly, Artogeia rapae, induced at least five groups of proteins with the antibacterial activity against certain Gram-negative and/or Gram-positive bacteria in the haemolymph. These antibacterial proteins were separated and one was purified by different types of chromatography. The purified protein was heat-stable and basic peptide, and its molecular weight was approximately 4 kDa. We propose the name hinnavins for this antibacterial peptide.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.307-316
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• 1996

A persistent major haemolyruph protein (MHP) was confirmed, and its ontogeny and physicochemical charadedstics were investigated in Helicoverpa assulta. The MHP existed continually during larval-pupal-adult development, and its ontogeny was similar to that of total haemolymph protein concentration during development. Its content increased with larval growth, and kept to high level during pupal-adult development except for temporary decrease at the early pupal and adult stages. The MHP was purified by ammonium sulfate precipitation, gel filtration and ion exchange chromatography. The purified MHP was determined to be hexamer glycolipoprotein (pI 5.9, M.W. 414kDa) consisted of single type subunit (69kDa). Amino acid analysis suggested that the MHP contained a relatively high content of aromatic amino acids (18.27 mole % of tryptophan, 7.47 mole % of tyrosine and 6.51 mole % of phenylalanine) compared to storage proteins from other insects. Immunodiffusion test and electrophoretic analysis of the organ proteins (gut, fat body, and Malphigian tubule) suggested that the major haemolymph protein was present in the fat body.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.75-80
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• 2006

Changes in the levels of protein and free amino acids in the haemolymph of three selected races of the silkworm, Bombyx mori viz., PM, $NB_4D_2$ and $CSR_2$, were investigated during 4th moult, 5th instar and pupal period. The levels of total protein in the haemolymph, increased from first day of 5th instar till sixth day. From seventh day till spinning, the protein levels decreased in all the three races. A sustained decrease in the haemolymph proteins was observed during the pupal development in all the three races. The levels of free amino acids, which were high during 4th moult, declined through the 5th age of larval development till spinning. PM showed a relatively higher free amino acid level (3.192 mg/ml) in haemolymph followed by $NB_4D_2$ (2.601 mg/ml) and $CSR_2$ (2.35 mg/ml). The free amino acid levels decreased gradually from prepupal stage but increased again at the end of pupal period. Racial differences in the changes in the levels of protein and free amino acids in the haemolymph were observed in the larvae and pupae when subjected to two high temperature regimes of $30^{\circ}C$ and $35^{\circ}C$. The results showed that high temperature induces specific changes in the metabolism (reversible thermal stress) that have different adaptive value in different races of the silkworm. Relatively higher increase in the free amino acid levels in the haemolymph of Pure Mysore presumably provides protective cover to tissues against high temperature by an increase in osmolarity and reduction in evaporative water loss. The absence of such a mechanism may be responsible for temperature susceptibility of the bivoltine races like $NB_4D_2$ and $CSR_2$.

• Journal of Sericultural and Entomological Science
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• v.27 no.1
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• pp.37-41
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• 1985

The haemolymph protein, digestive fluid proteins and digestive fluid amylase activity of wild silkworm, Theophila mandarina those of the were studied by polyacrylamide gel electrophoresis. In addition, they was also compared with silkworm. 1. 6 main protein bands in female and 7 main protein bands in male were detected in the larval haemolymph of T. mandarina where as 8 and 7 main protein bands in female and male of B. mori were observed. Some differences in the haemolymph protein ands of T. mandarina and B. mori were observed. 2. 15 protein bands and 12 protein bands were found in the larval digestive fluid of T. mandarina and B. mori respectively. Some differences in the mobility of digestive fluid proteins of T. mandarina and B. mori were noticed. 3. Larval digestive fluid amylases were anionic and moved near the tracking dye in both T. mandarina and B. mori. Mobility of the digestive fluid amylases relative to bromophenol blue were 0.019 and 0.020 in T. mandarina and B. mori respectively.

• The Korean Journal of Zoology
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• v.29 no.1
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• pp.1-12
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• 1986

Electrophoretic, immunological, and column chromatography methods were used to determine the appearance and distribution of storage proteins in various organs during the metamorphosis of Bombyx mori L. Two storage proteins start to appear in haemolymph in early 5th instar stage and show the identical mobility with fat body proteins. These proteins show the high concentration in haemolymph in last instar stage but accumulate in fat body after pupation. Storage protein-2 shows the distinct pattern for general storage proteins in both male and females. This protein is involved with the formation of cuticle protein in late last instar stage and appears to be temperally deposited in midgut during the pupal stage. Also SP-2 shows the identity with vitellogenin electrophoretically and immunologically and especially the positive reaction with antibody against yolk protein during the pupal stage, demonstrating that the storage protein is closely related to the formation of yolk protein.