SDS-Polyacrylamide gel 電氣泳動에 의한 누에의 發育段階別 血蛋白質의 變化

孫興大 건국대학교 1986 論文集 Vol.22 No.1

This experiments were carried out to study changes of haemolymph proteins during development of silkworm, Bombyx mori by SDS-polyacrylamide gel electrophoresis. The results obtained were follows; 1. Bands of haemolymph proteins seperated during the metamorphosis of silkworm showed 20 bands and those molecular weights ranged from 12,800 to 106,000 daltons. 2. Major bands (7 and 15) were changed quantitatively, band 7 showed predominant levels from the middle 5th instar to the middle pupa stage, band 15 was leached the maximum levels at spining and then significantly decreased on prepupa stage, those molecular weights were determined to be 27,000 and 79,000 daltons respectively. 3. Female specific proteins of band 17 and 18 (mol. wt. 102,000 and 102,500 daltons) showed from the 1st day after pupation. 4. Concentration of low molecular weight proteins (mol. wt. 16,300∼18,500 daltons) at adult stage were increased. 5. In conclusion, it was suggested that the major haemolymph proteins in silkworm should be utilized to form tissues of pupa and adult, female specific proteins involved in the egg formation.

들민달팽이(Deroveras varians)의 배자발육과 난황단백질

진병래,손흥대,박혜진,조은숙 東亞大學校附設 農業生命科學硏究所 1999 農業生命資援硏究 Vol.8 No.1

들민달팽이의 배자발육은 해부현미경으로 관찰하였다. 산란 후 4일이 경과면서 발육이 급격하게 진전되어, 10일이 지나면서 난 내에서 들민달팽이 유충의 형태를 갖추고, 12일째 부화하였다. 들민달팽이의 배자발육에 따른 난황단백질의 변화는 산란 1일째, 3일째 및 6일째 알의 난황단백질을 전기영동방법으로 분석하였다. 그 결과 배자발육과 함께 급격히 감소되는 난황단백질 관련 밴드를 관찰할 수 있었다. We characterized embryogenesis and egg proteins of Variable field Slug, Deroceras varians. The embryogenesis was observed by light microscopy. The eggs were hatched at the twelfth day after oviposition. The egg proteins of the first day, third day and sixth day after oviposition were analyzed by native-and SDS-PAGE analyses. The results showed that yolk proteins are gradually decreased during embryogenesis.

누에 배자와 난소의 초대 배양에 있어서 세포의 분화형태

최광호,손홍대,조은숙 東亞大學校生命資源科學大學附設 農業資源硏究所 1997 農業生命資援硏究 Vol.6 No.1

곤충 세포의 초대 배양에 있어서 그 분화양상을 관찰하기 위하여 누에 배자와 난소를 초대 배양하였다. 그 결과 몇 가지 상이한 세포형이 관찰되었는데 배자 초대 배양의 분화형태는 상피형 세포와 망상조직을 형성하는 섬유상 세포가 관찰되었고, 난소에서는 주로 상피형 세포가 관찰되었다. 또한 두 조직 모두에서 거대 세포보다는 작지만 다소 큰 세포 형태가 관찰되었다. This study was carried out to investigation for differential charaterization of the cells in primary culture with insects. In the process of primary culture with embryos and larval ovaries of silkworm, Bombyx mori, three types of cells were observed. Fibroblast-like cells which formed network and epitherial-like cells migrated from embryos. But in case of ovaries, fibroblast-like cells were not found. Somewhat large cells but smaller than giant-cells were observed in the cultured embryos and ovaries.

Production of the Eggs with Abnormal Shape from the Domestic Silkworm , Bombyx mori , Infected with Autographa californica Nuclear Polyhedrosis Virus

(Hung Dae Sohn),(Byung Rae Jin),(Sang Mong Lee),(Nam Sook Park),(Hye Jin Park),(Eun Young Yun),(Seok Woo Kang),(Keun Young Kim) 한국잠사학회 2000 International Journal of Industrial Entomology Vol.1 No.2

Molecular Cloning and Characterization of a Ferritin Heavy Chain Subunit of the Cricket Teleogryllus emma

Mi Ri Sohn,Nam Jung Kim,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

We describe here the cloning and characterization of a cDNA encoding the ferritin heavy chain homologue (TeFerHCH) from the cricket Teleogryllus emma. The TeFerHCH gene spans 1,009 bp and consisted of four introns and five exons coding for 217 amino acids residues. The TeFerHCH subunit contained the conserved motifs for the ferroxidase center typical of vertebrate ferritin heavy chains and the iron-responsive element (IRE) sequence with a predicted stem-loop structure was present in the 5'-untranslated region (UTR) of TeFerHCH mRNA. TeFerHCH was grouped with the S type (HCH) in a phylogenetic tree. The TeFerHCH cDNA was expressed as approximately 27 kDa polypeptide in baculovirus-infected insect Sf9 cells. Northern blot analysis revealed that TeFerHCH exhibited ubiquitous expression and was upregulated by wounding and iron overload in the fatbody, suggesting a functional role for TeFerHCH in iron metabolism.

Construction of Stably Transformed Bm 5 Cells by Using Autographa californica Nuclear Polyhedrosis Virus IE 0 Gene

Cho, Eun Sook,Jin, Byung Rae,Sohn, Hung Dae,Choi, Kwang Ho,Kim, Soung Ryul,Kang, Seok Woo,Yun, Eun Young,Kim, Sang Hyun,Kim, Keun Young,Je, Yeon Ho,Kang, Seok Kwon 한국잠사학회 1998 한국잠사곤충학회지 Vol.40 No.2

To construct transfurmed Bm5 cells, Autographa californica nuclear polyhedrosis virus(AcNPV) IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% nucleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/ml G4l8 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistant cells was isolated and characterized by PCR using AcNPV IEI gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. mori-derived Bm5 cells as well as Spodoptera fugjprrda-derived Sf9 cells to produce stably-transformed insect cells

Molecular Cloning and Characterization of a Phospholipase A2 of the Bumblebee Bombus ignitus

Yu Xin,Young Moo Choo,Hyung Joo Yoon,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

Among bee venom proteins, phospholipase A2 (PLA2) is critical one of bee venom components to defend against predators intruders. In this study, PLA2 gene from cDNA libarary using the venom glands of Bombus ignitus worker bees(BiVn-PLA2) was cloned and characterized. BiVn-PLA2 spans 2211 bp and consists of three introns and four exons encoding 180 amino acid residues. BiVn-PLA2 shares high levels of identity with a bumblebee, B. terristris (89% protein sequence identity), B. pennsylvanicus (88%), and a honey bee, Apis mellifera (53%). Northern blot analysis revealed that BiVn-PLA2 is expressed in venom gland, indicating that BiVn-PLA2 is one of the venom components of B. ignitus. To determine BiVn-PLA2 of venom components from venom sac, N-terminal amino acid sequencing of a putative BiVn-PLA2 (the purified 18 kDa) was performed by Edman degradation. The N-terminal amino acid sequencing of the 18 kDa protein was coincident with the N-terminal amino acid residues of the mature BiVn-PLA2 and the 18 kDa protein catalysed the hydrolysis of DBPC subs trate[1-O-(6-Dabcyl-aminohexanoyl)-2-O-(12-(5-B ODIPY-entanoyl) aminododecanoyl)-sn-glyceryl phosphatidylcholine] that is a sensitive fluorogenic probe for PLA2 activation. Western blot analysis revealed that BiVn-PLA2 is expressed in the venom gland, stored in the venom sac, and then emitted throughout sting apparatus. Finally, to test BiVn-PLA2 toxicity, BiVn-PLA2 was adjusted to a insect cell (Sf9) at different concentrations (1-30 μg/2×105 cells). The apoptotic cell death assay results showed that the cell survival decreased with increasing concentrations (1-30 μg/2×105 cells).

누에 體液의 幼若호르몬 結合蛋白質(Juvenile hormone binding protein)에 관한 硏究

孫興大 한국잠사학회 1988 한국잠사곤충학회지 Vol.30 No.1

In order to examine a physiological role of juvenile(JH) binding proteins in the hemelymph of the silkworm larva, Bombyx mori, [3H] JH I incubated hemolymph was separated by polyacrylamide gel electrophoresis in the fifth-instar larva and the activity of the binding protein was analyzed using charcoal binding assay. The results obtained were as follows; 1. The JH was bound by two protein fractions in the hemolymph of the fifth-instar larva; One was JH binding lipoprotein(JH-LP), thr other was JH specific binding protein (JHBP). Their relative mobility values (Rm) were 0.3~0.33 and 0.81~0.84, respectively. There were no valid differences in those values from developmental stages of both male and female silltworms. 2. Total protein contents of the hemolymph were gradually increased during the fifth-instar larva, while at the prepupa decreasd. The maximum ones were observed at the spinning period and the contents from female were much higher than those from the male. 3. JH binding activity per ml of the hemolymph was low in the early stage of the fifth-instar larva and its activity was maximized at the spinning period and at the prepupa slightly decreased. 4. There was a similar pattern between changes of the JH binding activity per ml of the homolymph and of the total protein contents of the hemolymph. 5. The JH binding activity per mg of the hemolymph proteins was high in the early stage of the fifth-instar larva, while from the 6th day of the fifth-instar larva to the prepupa its activity showed the lowest levels.

누에 유충의 혈림프 유약호르몬 에스테라제 활성의 조절에 關한 硏究

孫興大,姜弼敦 한국잠사학회 1992 한국잠사곤충학회지 Vol.34 No.1

Effects of starvation, ligation, refeeding and methoprene treatment in the feeding phase of the fifth instar lavae of Bombyx mori on the regulation of juvenile hormone esterase(JHE) activity were investigated. Starvation and ligation contributed to the reduction of JHE activity, however. JHE levels in starved lavae were slightly higher than in ligated larvae. Haemolymph JHE activity of starved larvae was increased by refeeding, and duration of increasing time of JHE activity after starvation was related to duration of starvation. When starved larvae were applied methoprene topically. JHE activity were not changed at day 0 and 1, but were increased by 1.3~1.4 times between day 2 and 5. When ligated larvae were applied methoprene topically, JHE activity was not changed at day 0, but were increased by 1.9~2.3 times between day 1 and 5. These results suggest that head factor, juvenile hormone(JH) and nutrient are major factors in the regulation of JHE in the feeding phase of the fifth instar larvae of Bombyx mori. Especially JHE might be regulated by the co-operative action of head factor and JH. However, head factor plays important role in the early stage, while JH plays important role thereafter.

幼若홀몬이 家蠶의 體液蛋白質 및 아미노酸의 變動에 미치는 影響

孫興大 한국잠사학회 1986 한국잠사곤충학회지 Vol.28 No.1

The experiment was carried out to investigate the effects of juvenile hormone analogue on changes of protein and amino acids in haemolymph of silkworm larva. Juvenile hormone analogue was topically administered to larvae at dose of 1 μg and 10μg Per gm of body weight at 60hr. of the 5th instar. The results obtained were as follows; 1. Larval duration of the fifth instar was extended about 1 day by JHA-1 and 4 days by JHA-10εs compared with the control. 2. Cocoon weight and cocoon layer weight by topical application of JHA were heavier than those of the control, but cocoon layer ratio was decreased in JHA-10 to the exclusion of JHA-1. 3. The concentration of haemolymph protein during the fifth instar was increased remarkably by the JHA application. 4. The total content of amino acids in haemolymph proteins of JHA-10 approximately doubled that of the control, with the conspicuous increase of glycine and arginine level.