• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.225-232
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• 1998

We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

• Journal of fish pathology
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• v.21 no.1
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• pp.67-79
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• 2008

This study was conducted to find out biological response of bivalves exposed to tributyltin chloride(TBTCl). The results of the study confirmed that TBTCl induce the reduction of oxygen consumption rateand histopathological feature in the gill structure of equilateral venus, Gomphina veneriformis. The experi-mental groups consisted of a control and 3 TBTCl exposure groups (0.4, 0.6 and 0.8 yg TBTCl L') and theexperimental period was 36 weeks. For histological analysis, gill tissues were fixed in Bouin's fluid andthen stained H-E stain, AB-PAS (PH 2.5) reaction and Masson's trichrome stain after having serially sec-tioned the tissue by paraffin method at thickness of 4-6 (an. The oxygen consumption rate was not signifi-cantly different between the control and exposure groups at 4 weeks, but in all exposure groups at 28 weeks,it was significantly different to the control. Gill of G. veneriformis had demibranch that attached two sheetsof lamellae and a lamella was composed of numerous filaments, numbering 25 on average. The frontal fila-ment zone had three types of cilia; frontal, latero-frontal and lateral depending on locations while the lateralcilia were the longest and largest in number. The mucous cells observed in filaments were more abundant in(542c) in AB-PAS (PH 2.5) reaction. Gill exposed to TBTCl was extended hemolymph sinus and increased hemocytes at 4 weeks, and then it showed increases of mucous cells and partially disappearance of frontalcilia. In the group of 0.8 yg TBTCl L' at 12 weeks, hypertrophy of frontal and latero-frontal epithelia wasobserved. Also it observed m decrease of mucous cell containing weekly acid mucosubstance and appearedpartially destruction muscle fiber bundle, In the groups of 0.4 and 0.6 ug TBTCl L' at 36 weeks, it appearedpartially modification of epithelia and in 0.8 us TBTCl L' group, observed filaments that come out chiti-nous rod from disappearance of frontal and latero-frontal epithelia.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.161-166
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• 2001

The storage proteins of the mulberry longicorn beetle, Apriona germari Hope, were purified and characterized. Three kinds of storage protein (SP1, SP2 and Sp3) were purified from the last instar larval hemolymph of A. germari by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. The SP1, SP2 and SP3 have a native molecular weight of 480, 440 and 420 kDa, respectively. In the SDS-polyacrylamide gel electrophoresis analysis, these storage proteins are composed of a single protein subunit with molecular weight of 90, 85 and 80 kDa, respectively. This result showed that the storage proteins are hexameric protein. The SP1 and SP2 were stained with Schiffs reagent, but SP3 was not stained. It can be assumed that SP1 and SP2 are glycoprotein. Western blot analyses using the each of polyclonal antiserum against purified SP1, SP2 and SP3 showed that the three antibodies reacted with the each of SP bands, respectively. Also, antibodies against SP1 and SP3 cross-reacted with the SP3 and SP1, respectively. However, SP2 was not cross-reacted with these two antibodies. Also, antiserum against SP2 did not cross-reacted with the SP1 and SP3.

• Applied Microscopy
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• v.35 no.3
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• pp.177-186
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• 2005

The foregut epithelium of the last instar larva in the scarab beetle, Allomyrina dichotoma was observed with electron microscopes. The foregut epithelium of the scarab beetle larva is composed of a single-layered squamous absorptive cell. The luminal surface of the epithelium is covered with cuticular intima. The free surface of the squamous cell has a irregular array of microvilli 'brush border', while cell membrans close to the basal lamina are infolded and a lot of mitochondria are concentrated in those processes. The cytoplasm in the epithelial cells is well developed nucleus, mitochondria. And the basal region of cell contains large lipid-, protein droplets and numerous glycogen granules. The basal lamina is located between the basal membrane and muscle bundle, providing barrier between the epithelium and the hemolymph. The epithelium is surround by the subepithelial space and muscles. The subepithelial space, which is composed of fibrous connective tissue is innervated by many tracheoles and axon.

• Journal of Life Science
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• v.9 no.1
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• pp.84-89
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• 1999

To determine phylogenetic relatedness of four silk-producing silkmoths (B. mori, B. mandarina, A. yamamai and A. pernyi), internal coding region of arylphorin which is a storage protein in hemolymph protein of insects were amplified by polymerase chain reaction and then sequenced and compared each other. The nucleotide composition was biased toward adenine and thymine(59% A+T) and a strong bias for use of C in the third position of codons was found for Phe and Tyr. Together TTC(Phe) and TAC(Tyr) account for about 16.8% (10 for TTC and 8 for TAC) of all codon usage. The nucleotide similarity of arylphorin gene from B. mori showed 99%, 98% and 97% homology with those of B. mandarina, A. yamamai and A. pernyi, respectively. Also, the nucleotide sequence of arylphorin gene from B. mandarina showed 98% and 97% homology with those of A. yamamai and A.pernyi, respectively. Between A. yamamai and A. pernyi, the sequence homology was 97%. The deduced amino acid sequences in B. mori, B. mandarina and A. yamamai showed almost 99% homology. Although the aryphorin gene provided insufficient variability among the four insect species, A UPGMA tree is generated that supported the monophyly of silk-producing insects, with M. sexta placed basal to it. It is suggest that silk-producing insects have a close relationship and a homogeneous genetic background from comparison with those of other insects.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.269-275
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• 1998

Spider crabs, Libinia emarginata, were eyestalk-ablated unilaterally and bilaterally to manipulate endogenous methyl farnesoate (MF) to increase during the embryogenesis. Endogenous MF were measured weekly over the embryogenesis of the crab, using HPLC with the aids of GC/MS and MS database (CAS 010485-70-8) for the identification of the hormone. Initial MF titers both in the hemolymph and embryos of intact control were at bottom levels and the hormone concentrations kept unchanged (p<0.05), reflecting physiological unnecessaries of the hormone in the embryogenesis. Eyestalk ablation significantly stimulated the crabs to increase endogenous MF in both tissues (p<0.0l). In the response of the embryos to the increased MF, no growth stimulations were observed, at least, in the first part of embryogenesis. The increased mortalities and immature sheddings of embryos resulted from the crabs under the influence of elevated MF in both tissues, instead, suggesting that the elevated MF against the crab's requirement blocked the normal developmental process of the crab embryos. These data can give crustacean endocrinologists some insights to understand the effects of the hormone on the crustacean reproduction studied previously in which JH analogs ambiguously affected the crustacean reproduction depending on the reproductive stages. The data also can give shrimp aquaculturists some implication of a possible generation of unfavorable shrimp seeds attributed to elevated egg MF originated from their eyestalk-ablated mother shrimp.

• BMB Reports
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• v.46 no.7
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• pp.358-363
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• 2013

In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti-Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein.

• The Korean Journal of Malacology
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• v.31 no.2
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• pp.151-158
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• 2015

This study was conducted to find out the changes of survival, respiration and organ structure of Tegillarca granosa exposed to copper (Cu). Experimental period was four weeks. Experimental groups were composed of one control condition and three copper exposure conditions (0.125, 0.250 and 0.500 mg/L). The results of the study confirmed that copper induces reduction of survival rate and respiration rate and histopathology of organ structure of the bivalve. In the copper concentration of 0.500 mg/L, mortality was 66.7% after Cu exposure of 4 weeks. Respiration rate was observed exposure groups lower than control decline by 18%. Histological analysis of organ system illustrated degeneration of epithelial layer and connective tissue layer of the mantle. Also, histological degenerations as epithelial atrophy and disappearance of lateral cilia are recognized in the gill and it was observed expansion of hemolymph sinus, disruption of epithelial layer, acidification of mucous and degeneration of muscle fiber bundles in the foot. In the digestive diverticulum, it was showed atrophy and destruction of basophilic cell and epithelial cell in the digestive tubules.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.145-149
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• 2003

ApolipophorinIII (apoLp-III) is a protypical exchangeable apolipoprotein that is abundant in hemolymph of many insect species. Its function lies in the stabilization of low-density lipophorin particles (LDLp) crossing the hemocoel in phases of high energy consumption to deliver lipids from the fat body to the flight muscle cells. But, recent studies with naive Galleria mellonella-apoLp-III gave first indication of an unexpected role of that protein in insect immune activation. In this research, we cloned a cDNA encoding putative apoLp-III from the silkworm, Bombyx mori injected with E. coli and characterized its role. We constructed a cDNA library using whole bodies of B. mori larvae injected with E. coli, carried out the differential screening, and selected the up-regulated clones. Among these clones, we focused on a cDNA showing a high sequence similarity to the apolipophorinIII from other insects and analyzed the nucleotide and deduced amino acid sequences. The pupative B. mori Jam123 apoLp-III cDNA contained 1,131 bp encoding 186 amino acid residues. Phylogenetic analysis revealed that the nucleotide and amino acid sequences of the B. mori apoLp-III cDNA formed a highly inclusive subgroup with Bombycidae. But, it was interesting that B. mori Jam123 is closer to B. mandarina than B. mori P50 and B. mori N4. Northern blot analysis showed a signal in the fat body, posterior silkgland and midgut.

• The Korean Journal of Malacology
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• v.28 no.1
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• pp.37-43
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• 2012

In this study, we invesgated a cytogenetic analysis of the Pacific triploid abalone, Haliotis discus hannai induced by low temperature treatment. We got a lot of mitotic metaphase chromosome spreads from the triploid and diploid Pacific abalones' hatched larvae (trochophores). The chromosome number of diploid abalone was 2n = 36 and that of triploid abalone was 3n = 54, so the chromosome number of triploid abalone was 1.5 times higher than that of diploid abalone. We developed a modified flow cytometric method for Pacific abalone from the existing methods. We uesd 51 months aged triploid and diploid Pacific abalones' hemolymph to get the DNA contents by flow cytometry. The DNA content of diploid abalone was 1.743 pg/cell and the DNA content of triploid abalone was 1.49 times higher than that of diploid one. It proved that triploid abalone was consisted with two sets of maternal diploid and one set of paternal genome.