• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.2
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• pp.37-48
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• 2008

The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.1
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• pp.31-40
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• 2013

Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.117-121
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• 2004

The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.95-96
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• 2003

We investigated the proteins that are associated with initiation, maintenance, and termination of diapause in silkworm eggs by means of 2D-gel electrophoresis. 1. Materials : Bombyx mori: unfertilized eggs, diapause eggs, non-diapause eggs, cold-treated egges. 2. Methods: isoelectric focusing, SDS-PAGE, silver staining. (omitted)

• Korean journal of applied entomology
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• v.27 no.1
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• pp.6-13
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• 1988

Studies were carried out to investigate the diapause-sensitive stage, the effect of 20-hydroxye- cdysone on diapausing pupae and the changes in pupal proteins of the fitll webworm, HYPha¬ntria cunea Drury. The stage sensitive to diapause-inducing photoperiod lies between 5th and 15th day of the larval development. And the oxygen consumption rate of diapausebound pupae decreased to a mean level of $30mu$l $O_{2}$/g/hr. The 20-hydroxyecdysone was effective in raising the respiratory acivity of 2.5% homogenate of the diapausing fall webworm pupae and in causing a normal adult development of the diapause-bound pupae. The soluble protein content rapidly declined in normally-developing pupae, but in diapause-bound pupae it was more or less static with much higher quantity. And protein bands separated from fat body of diapause¬bound pupae were different from those of nondiapausing pupae.

• Korean journal of applied entomology
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• v.34 no.4
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• pp.287-294
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• 1995

Studies were carried out to elucidate the efficacy of exogeneous 20-hydroxyecdysone treatment on terminating pupal diapause in the fall webworm. Hyphantria cunea Drury. And the difference was also investigated between normal adult development and post-diapause development after 20-hydroxyecdysone treatment. In the diapause termination rate of pupae treated with 20-hydroxyecdysone after storage for various periods at 16L:8D and $25\pm$$1^{\circ}C$, at the highest rate was observed from the group stored for the longest period and the lowest rate from those stored for 1.5 months. The time needed for adult emergence was inversely proportional to the chilling (at $0\pm$$1^{\circ}C$) period, and the longer its exposure period at low temperature, the higher its sensitivity to 20-hydroxyecdysone treatment. Pupal diapause duration was almost the same, regardless of storage period in the total darkness or at the photoperiod of 16L:8D, and also they successfully emerged to adult even without any experience at low temperature. The oxygen consumption rate in normally developing pupae showed nearly a typical U-form. But, that of diapausing pupae treated with 20-hydroxyecdysone slowly increased and jumped 14 days after the treatment. Pupal diapause began before formation of adult tissues, an a timing of adult tissue formation coincided with ascending timing of the metabolic rate in both normally developing pupae and diapausing pupae treated with 20-hydroxyecdysone. The diapausing pupae treated with 20-hydroxyecdysone was similar to normally developing pupae in band patterns of proteins from haemolymph or fat body.

• Korean journal of applied entomology
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• v.32 no.2
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• pp.139-150
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• 1993

This study was conducted to investigate the effect of 20-hydroxyecdysone on diapausing pupae of Hyphantna cunea. Treatment of 20-HE at the dosage of 5 $\mu$ g or higher/g live weight terminated diapause in about 50% of Hyphantna cunea pupae treated, and some malformed adult emerged at dosages of 10 or 12.5 $\mu$g of 20-hydroxyecdysone/g live wt. The maximum concentration of 20-hydroxyecdysone in the whole body of normally developing pupae was about 3.2 $\mu$g/g on the 6th day after pupation, and in that of diapausing pupae treated with 20 -HE about 4.1 $\mu$g/g on the 6th day after the treatment. But diapausing pupae showed a low level( 1.7 $\mu$g/g) of maximum 20-HE concentration. In diapausing pupae treated with 20- HE, glycongen content was lower than in normally developing pupae, but the changing pattern was similar to that observed in normally developing pupae. Glucose(and/or sorbitol) and trehalose content of 20-HE-injected pupae reached the maximum value on the 6th day after the injection, which were higher than those of normally developing pupae. The amount of soluble haemolymph proteins was lower but the total soluble protein content of haemolymph-rernoved whole body was higher in 20-HE-injected pupae than that in normally developing pupae.

• BMB Reports
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• v.39 no.6
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• pp.749-758
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• 2006

The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the full-length cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heat-stresses. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase.