• International Journal of Industrial Entomology and Biomaterials
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• v.14 no.1
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• pp.51-56
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• 2007

In a previous study, three larval cuticle protein genes were cloned from the mulberry longicorn beetle, Apriona germari (Comp. Biochem. Physiol. B 136, 803-811, 2003). In the present study, the genomic structures of these three larval cuticle protein genes (AgLCP9.2, AgLCP12.6 and AgLCP12.3) were elucidated. All three cuticle protein genes consist of one intron and two exons. Southern blot analysis of genomic DNA suggested that three cuticle protein genes are a single copy gene. In addition, a novel larval cuticle protein gene, AgLCP10.6, was cloned from A. germari in this study. The AgLCP10.6 cDNA contains an ORF of 300 nucleotides that are capable of encoding a 100-amino acid polypeptide with a predicted molecular mass of 10.6 kDa. The amino acid sequence deduced from the AgLCP10.6 cDNA contained a type-specific consensus sequence identifiable in other insect cuticle proteins and is most homologous to Drosophila melanogaster cuticle protein ACP65A (51 % protein sequence identity). Northern blot analysis revealed that AgLCP10.6 showed epidermis-specific expression.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.275-286
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• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Total weight gain, daily feed intake and feed conversion rate for 10 days were significantly improved to 56%, 20% and 22% in the piglets fed plasma protein, respectively. 2. A significant increase in N (null or non T/non B) cells was also noticed. Leukocyte proportion from piglets fed plasma protein was 20.2-24.7%, otherwise that from piglets fed without plasma protein was 12.3-13.4%, respectively. 3. A significant increase in the proportion of B cells and cells expressing poCD1 was not found in piglets fed plasma protein. 4. Reaction with monoclonal antibodies specific to adhesion molecules, poCD11a, poCD11b, poCD44 and poCD45A and poCD45B, has shown that leukocyte subpopulation from piglets fed plasma protein did not significantly higher than that from piglets fed without plasma protein. 5. Total proportion of granulocytes and monocytes was about 50% in both group and the proportion after treated with Hypaque/Ficoll was 2.7% and 5.8% in each group, respectively.

• Journal of the Korean Society of Food Culture
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• v.5 no.4
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• pp.443-448
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• 1990

The object of this study was to investigate the characteristics of pectinesterase(PE), polygalacturonase(PG) and Peroxidase(POD) in Kimchi materials. The results were as follows : 1. The specific activities of PEs in Korean cabbage, Korean raddish, garlic and ginger were 200 unit/mg protein, 23.1 unit/mg protein, 0.8 unit/mg protein and 32 unit/mg protein, respectively. The optimum pHs of PEs in all materials were between 7 to 8. The concentrations of NaCl, $CaCl_2$ which showed the highest activities of PEs were $0.2{\sim}0.3M$ NaCl, 50 mM $CaCl_2$ in Korean cabbage and raddish, 0.05 M NaCl, 20 mM $CaCl_2$ in garlic and 0.2 M NaCl, 20 mM $CaCl_2$ in ginger. 2. The specific activities and the optimum pHs of PGs were 1.5 unit/mg protein and pH 4.5 in Korean cabbage, 1.6 unit/mg protein and $pH\;4.5{\sim}5.5M$ in Korean raddish, 0.06 unit/mg protein and $pH\;3.0{\sim}3.5M$ in garlic, and 0.06 unit/mg protein and $CaCl_2$ in ginger. The concentrations of NaCl, $CaCl_2$ which showed the highest activities of PGs were $0.1{\sim}0.2M$ NaCl and $0.15{\sim}0.2mM$ mM $CaCl_2$ in all materials. 3. The specific activities and the optimum pHs of PODs in Korean cabbage, Korean raddish, garlic and ginger were 71.3 unit/mg protein ; pH 6.0. 769 unit/mg protein ; pH 5.5, 1.09 unit/mg protein ; pH 4.5 and 12.7 unit/mg protein ; $pH\;5.0{\sim}5.5M$, respectively. POD activities were not decreased in Korean cabbage, but decreased in Korean raddish by the increase of NaCl, $CaCl_2$ concentrations. In garlic and ginger, POD activities were a little slightly affected by the increase of NaCl, $CaCl_2$ concentrations.

• KSBB Journal
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• v.10 no.4
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• pp.455-460
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• 1995

Specific growth rate was controlled for the repression of acetic acid formation in the fed-batch fermentation of recombinant Escherichia coli. With controlled specific growth rate, we studied the effect of the specific growth rate on cell growth, glucose consumption, acetic acid formation, and the expression of recombinant protein (${\beta}$-lactamase). High specific growth rate caused the accumulation of glucose and acetic acid, and lowered the production of recombinant protein. However, the addition of methionine recovered the gene expression by alleviating the negative effect of acetic acid at high specific growth rate.

• Journal of fish pathology
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• v.23 no.3
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• pp.273-280
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• 2010

To determine whether rock bream Oplegnathus fasciatus can be protected from rock bream iridovirus (RBIV) infection by intramuscular injection of long double-stranded RNAs (dsRNAs), we compared protective effect of virus-specific dsRNAs corresponding to major capsid protein (MCP), ORF 084, ORF 086 genes, and virus non-specific green fluorescent protein (GFP) gene. Furthermore, to determine whether the non-specific type I interferon (IFN) response was associated with protective effect, we estimated the activation of type I IFN response in fish using expression level of IFN inducible Mx gene as a marker. As a result, mortality of fish injected with dsRNAs and challenged with RBIV was delayed for a few days when comparing with PBS injected control group. However, virus-specific dsRNA injected groups exhibited no significant differences in survival period when compared to the GFP dsRNA injected group. Semi-quantitative analysis indicated that the degree of antiviral response via type I IFN response is supposedly equal among dsRNA injected fish. These results suggest that type I IFN response rather than sequence-specific RNA interference might involve in the lengthened survival period of fish injected with virus-specific dsRNAs.

• IMMUNE NETWORK
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• v.11 no.3
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• pp.182-189
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• 2011

Background: Cytotoxic T lymphocytes (CTLs) appear to play an important role in the control and prevention of human cytomegalovirus (HCMV) infection. The pp65 antigen is a structural protein, which has been defined as a potential target for effective immunity against HCMV infection. Incorporation of an 11 amino acid region of the HIV TAT protein transduction domain (Tat) into protein facilitates rapid, efficient entry into cells. Methods: To establish a strategy for the generation of HCMV-specific CTLs in vitro, recombinant truncated N- and C-terminal pp65 protein (pp65 N&C) and N- and C-terminal pp65 protein fused with Tat (Tat/pp65 N&C) was produced in E.coli system. Peripheral blood mononuclear cells were stimulated with dendritic cells (DCs) pulsed with pp65 N&C or Tat/pp65 N&C protein and immune responses induced was examined using IFN-${\gamma}$ ELISPOT assay, cytotoxicity assay and tetramer staining. Results: DCs pulsed with Tat/pp65N&C protein could induce higher T-cell responses in vitro compared with pp65N&C. Moreover, the DCs pulsed with Tat/pp65 N&C could stimulate both of $CD8^+$ and $CD4^+$ T-cell responses. The T cells induced by DCs pulsed with Tat/pp65 N&C showed higher cytotoxicity than that of pp65-pulsed DCs against autologous lymphoblastoid B-cell line (LCL) expressing the HCMV-pp65 antigen. Conclusion: Our results suggest that DCs pulsed with Tat/pp65 N&C protein effectively induced pp65-specific CTL in vitro. Tat fusion recombinant protein may be useful for the development of adoptive T-cell immunotherapy and DC-based vaccines.

• Korean Journal of Biological Psychiatry
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• v.3 no.2
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• pp.170-180
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• 1996

Lowered immune function in the senile dementia patients may be related to the abnormal metabolism of amyloid precursor protein(APP). To investigate the passibility of an abnormal metabolism of APP in lymphocytes and the possible role of APP in the activation of lymphocytes in senile dementia patients, immunohistochemical study of rat spleen and fluorescence activated cell sorter analysis(FACS) of human lymphocytes with the specific antigen far each lymphocyte and double fluorescent marker with antibody to APP were performed. After stimulating lymphocyte with phytohemagglutinin(PHA), APP mRNA and protein were extracted and quantitfied and the influence of ${\beta}$-amyloid protein($A{\beta}$) specific antibody on lymphocyte division was investigated. In spleen, the majority of cells showing $A{\beta}$ immunoreactivity was found in the T-sell dependent zone. FACS indicated that around 90% $CD_4(+)$ T-cells and 60% of $CD_8(+)$ T-sell were immunoreactive to $A{\beta}$ specific antibody(mAb 4G8). Northern blot analysis shows that lymphocyte APP mRNA was gradually increased to reach a maximum at 3 days after activation with lectin mitogen PHA. However, the $A{\beta}$ immunoreactivity an cell surface remained constant during stimulation with PHA, indicating that the release of APP(secreted farm of APP) might be increased. A very large increase in soluble APP secretion was observed in T-lymphocyte upon activation, but only law levels in the resting stale. Immunoblot was carried out an the protein obtained from cell lysate after stimulating lymphocyte by applying PHA to the cultured lymphocyte, and the result was that $A{\beta}$ band of immature farm under 116 KDa marker decreased as the duration of culture was increased after PHA stimulation. The monoclonal $A{\beta}$ specific(4G8) and polyclonal APP antibodies did not inhibit the [$^3H$]-thymidine uptake of mitogen-treated lymphocytes significantly, suggesting that mitogenesis can not be inhibited by specific $A{\beta}$ and polyclonal APP antibody. These results suggest that APP is expressed in T-cell and might be closely associated with the function of T-cells.

• Journal of Microbiology
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• v.33 no.2
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• pp.128-131
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• 1995

Using differential hybridization, we selected the prk gene fortuitously from Schizosaccharomyces pombe homologous to RACK1 of rat which encodes the receptor for activated protein kinase C. The cDNA sequence of prk was determined and its deduced amino acid sequence was 76% homologous to RACK1 and had the feature of trimeric G protein bata subunit. The specific amino acid sequences required for the protein kinase C binding were also present in Prk as in the case of RACK1 protein. From these similarities, we suggest that the Prk is protein kinase C binding protein of S. prombe. The involvement of Prk in signal transduction mediated by protein kinase C remained to be studied.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.269-276
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• 1993

To ascertain that tegument of Pnragonimus westermoni has specific antigenic proteins, the tegumental fraction was isolated from 10-month-old worms by 0.1% digitonin solution, and subjected to SDS-PAGE and immunoblot. Component proteins of tegumental syncytium comprised of 94, 74 (76-66), 62, 54, 44, 42, 38, 28, 26, 25, 24, 17, 15.5 and 13.5 kDa proteins. Of them, the 94, 44 and 42kDa proteins were more specific to tegument, especially the 94 kDa protein was the most prevailing one. In immunoblot, antigens of the 94, 90, 78, 76, 74, 68, 65, 63, 60, 59 and 54 kDa proteins were commonly detected by 7 sera of 10 human paragonimlasis, but none of them reacted with 5 sera of clonorchiasis. In conclusion, the 94 kDa protein was the major tegumental protein, as well as the specific antigen. The 76 and 66 kDa proteins were the minor components of tegument, which were also specific antigens of R westermcni.

• BMB Reports
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• v.33 no.1
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• pp.92-95
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• 2000

The HIV-1 p24(202-221) sequence ETINNEEEWDRVHPV HAGP contains a B-cell epitope with the earliest immune response and the highest antibody titer against anti-mouse sera obtained by immunization with p24 antigens. A novel mouse monoclonal antibody (mAb) was generated against the immunodominant B-cell epitope of the HIV-1 p24 capsid protein, p24(202-221). BALB/c mice were immunized with the four branched multiple antigenic peptide (MAP) containing the HIV-1p24(202-221) sequence, and antibody-secreting hybridoma were produced by fusion of mouse splenocytes with P3X63Ag8.653, mouse myeloma cells. One clone which produced the antigen-specific mAb named KI-24 (Isotype IgG1, light chain: ${\kappa}$) was identified. mAb KI-24 was highly specific for both the p24(202-221) and p24 proteins when analyzed by ELISA and Western blotting. Since p24(202-221) also contains a cytotoxic T-lymphocyte epitope, this specfic peptide epitope and the monoclonal antibody with specific reactivity against the p24 protein and p24(202-221) can be used in peptide vaccine development and p24 antigen detection from HIV patients.