• International Journal of Industrial Entomology and Biomaterials
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• 제16권2호
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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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• 제26권1호
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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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• 제9권1호
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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.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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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)

• 한국응용곤충학회지
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• 제27권1호
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• pp.6-13
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• 1988

흰불나방 (Hyphantria cunea Drury) 의 용휴면이 결정되는 시기, 휴면용에서의 20-hydroxyecdysone 의 영향과 전기영동 방법으로 측정된 용단백질의 변화는 다음과 같았다. 용휴면의 결정은 유충발육기간중 초기 5~15일에 이루어졌다. 산소소비율의 변화는 정상발육용에서 U자 모양의 곡선을 나타내었고, 휴면용에서는 $30mu$l {TEX}$O_{2}${/TEX} /g/hr 수준으로 감소되어 유지되었다. 휴면용에서 추출된 2.5%균질액은 20-hydroxyecdysone 의 처리로 호흡활성이 증가하였고, 휴면예정용은 직접 주사로 우화하였다. 수용성 단백질의 함량 변화는 정상발육용에 급격하게 이루어졌으나, 휴면용에서는 완만하였고, 더 많은 양이 포함되어 있었다. 휴면예정용의 지방체에서 분리한 단백질의 종류는 비휴면용의 경우와 달랐다.

• 한국응용곤충학회지
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• 제34권4호
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• pp.287-294
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• 1995

미국흰불나방(Hyphantria cunea Drury) 번데기의 휴면 종료에 미치는 탈피호르몬 20-hydroxyecdysone (20-HE)의 처리효과와 또한 호르몬처리로 유기된 휴면 후 성충발육 모습을 정상적인 성충발육과 비교하기 위하여 본실험을 수행하였으며, 그 결과 다음고 같다. 광주기 16L.8D와 $25\pm1^{\circ}C$에서 여러가지 다른 기간동안 저장했던 휴면번데기에 20-HE를 처리했을 때 가장 높은 휴면 종료율은 가장 오랫동안 처리된 군에서 나타났고 45일 동안 저장했던 휴면번데기에서 가장 낮은 종료율을 보였다. 우화에 필요한 기간은 저온($0\pm1^{\circ}C$)처리가간에 반비례하였고, 또한 저온처리기간이 길어질 수록 20-HE에 대한 민감도는 높아졌다. 번데기 휴면기간은 그들이 16L.8D 광조건에 처리되었건 혹은 지속적이 암조건에 저장되었건 간에 거의 같았고, 또한 저온처리없이도 성공적으로 성충이 출현하였다. 정상적으로 무화(휴면없이)하는 번데기의 산소소모율은 전형적인 U자 모양을 보였지만 20-HE에 처리된 휴면 번데기의 산소흡수량은 천천히 증가하다가 처리 후 14일에 급격히 증가하였다. 번데기 휴면은 성충조직을 형성하기 이전에 들어갔으며, 정상적으로 발육하는 번데기나 또는 20-HE에 처리되어 발육하기 시작하는 휴면 번데기 모두에서 성충조직 형성시기와 대사율이 증가하기 시작하는 시기는 일치하였다. 탈피호르몬처리로 발육을 시작한 휴면 번데기의 혈림프와 지방체에서 분리된 단백질종류는 모두 정상적으로 발육하는 번데기의 조직에서와 같았다.

• 한국응용곤충학회지
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• 제32권2호
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• pp.139-150
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• 1993

미국흰불나방 휴면번데기에 5 $\mu\textrm{g}$/g(생체중) 이상의 탈피호르몬을 처리하면 처리된 용의 약 50%가 우화했으며, 10 $\mu\textrm{g}$이나 12.5 $\mu\textrm{g}$/g의 처리구에서는 몇마리의 기형적인 성충이 나타났다. 한편 탈피호르몬의 체내함량을 적정한 결과 정상용은 용화 후 6일째에 약 3.2 $\mu\textrm{g}$/g의 최대값을 보였으며, 탈피호르몬을 처리한 용에서는 처리 6일째에 4.1 $\mu\textrm{g}$/g의 최대값을 보였다. 그러나, 휴면 용에서는 낮은 탈피호르몬 함량을 보였는데 측정 기간중 얻은 최대치는 1.73 $\mu\textrm{g}$/g 정도였다. 탈피호르몬을 처리한 휴면 용의 글리코겐함량은 정상용 보다 낮았으나, 성충조직 발육기간 동안의 발육양상은 정상 용의 경우와 비슷하였다. 또한 같은 처리를 받은 휴면 용의 포도당-솔비톨과 트레할로스의 농도는 처리 후 6일째에 최대값을 보였는데, 그 수준은 정상적으로 발육하는 개체의 경우보다 높았다. 탈피호르몬 처리를 받은 휴면용의 혈림프 내 수용성단백질 함량은 정상 용의 그것보다 낮았으며, 혈림프를 제외한 전체 조직 내에서 정상용의 경우보다 높았다.

• BMB Reports
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• 제39권6호
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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.