• Animal cells and systems
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• v.2 no.1
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• pp.93-97
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• 1998

Coelomoic fluid protein (CP), a vitellogenin contained in the coelomic fluid of polychaetes, is transported by receptor-mediated endocvtosis that is controlled by GTP-binding proteins. Transport of 125l-CP was markedly inhibited by AlF4 and toxins such as cholera toxin and pertussis toxin. These effects appear to be mediated by cAMP, since 125l-CP transport was also greatly inhibited by dibutyryl cAMP. The results strongly suggest that hetero trimeric G-protein is involved in the regulation of 125l=CP transport through the activation of adenylyl cyclase. Immunoblotting tests with antibodies against Gsa and Gia subunits showed a Gsa subunit of 45 kDa in the membrane of oocytes of intermediate and large size classes and a Gia subunit of 41 kDa only in the oocytes of the intermediate size class.

• Biomedical Science Letters
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• v.18 no.3
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• pp.210-217
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• 2012

Oncolytic viral vectors have shown good candidates for cancer treatment but have many limitations. To improve the therapeutic potential of oncolytic vaccinia virus, we developed a recombinant vaccinia virus expressing the 4-1BBL co-stimulatory molecule or CCL21. 4-1BBL and CCL21 expression was identified by FACS analysis and immunoblotting. rV-4-1BBL vaccination shows significant tumor regression compared to rV-LacZ, but rV-CCL21 shows rapid tumor growth compared to rV-LacZ in the poorly immunogenic B16 murine melanoma model. 4-1BBL expression resulted in the increase of the number of CD8+ T cells and especially the increase of effector (CD62L-CD44+) CD8+ T cells. These data suggest 4-1BBL may be the potential target for enhancement of tumor immunotherapy.

• IMMUNE NETWORK
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• v.3 no.1
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• pp.23-28
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• 2003

Background: Intracellular antibody specific to hepatitis B virus X protein (HBx) might be useful for studying the role of HBx in hepatocellular carcinogenesis and HBV replication. Methods: With variable region genes for H7 monoclonal anti-HBx Ab, we constructed a vector for bacterial expression of single chain Ab (scFv) and a vector for eukaryotic cell expression of it. The expression of H7 scFv and its binding activity against HBx was examined by immunoblotting and immunofluorescence microscopy. Results: H7 scFv expressed in bacterial cells retained reactivity to HBx. We demonstrated its intracytoplasmic expression in CosM6 eukaryotic cells. Conclusion: This is the first study showing the expression of intracellular anti-HBx Ab in eukaryotic cells. H7 scFv may be a good tool to study the function of HBx in HBV infection.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.273-273
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• 2004

This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. (omitted)

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.193-199
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• 1993

The protein po;vmorphisms and allele frequencies of wild house rat (Rattus norvegicus) population in Korea were studied. The studied proteins and enzymes were transferrin (Tf), albumin (Alb), fumarate hvdratase (FH), phospho!loucomutase (PGM), lactate dehvdrogenase A (LDHA) and lactate dehvdrogenase B (LDHB). There were two transferrin alleles, TP and Tf in Korean wild house rat popu1ation. The Tf2 allele was found for the first time by a starch gel, and confirmed by a polvacrvlamide gel isoelectric focusing and immunoblotting. The allele frequencies of TP and TF were 0.985 and 0.015, respectively. Two common alleles fumarate hydratase, FHa and FHb were found, and frequencies of FHa and FPP were calculated to be 0.714 and 0.286, respectively. The kequenw of FH in Korean wild house rat was higher than that of Finnish and Czechoslovakian population. Alb, PGM, LDHA and LDHB are only one phenotype each and all. Therefore, these proteins seem to be monomorphic in Korean wild house rat population.

• BMB Reports
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• v.30 no.1
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• pp.80-84
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• 1997

The synthesis and secretion of human angiogenin in E. coli by the natural leader sequence has been studied. We constructed a recombinant plasmid containing human angiogenin cDNA which encompassed all the coding region including leader sequence required for secretion. The recombinant plasmid was introduced into a suitable E. coli host. The angiogenin was detected in the culture medium and periplasm upon the induction of gene expression. The molecular weight of the secreted angiogenin was identical to that of authentic angiogenin purfied from human plasma when estimated by SDS-PAGE and immunoblotting. showing that the natural leader sequence was recognized and processed by the secretion machinery of E. coli. The angiogenin concentration in the culture medium reached a maximum within 2 h when expressed at $37^{\circ}C$ with 0.02~2 mM IPTG. In contrast, the expression level increased gradually over time up to 11 h at $23^{\circ}C$ with 0.002~2 mM IPTG and at $37^{\circ}C$ with 0.002 mM IPTG.

• Microbiology and Biotechnology Letters
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• v.25 no.6
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• pp.612-616
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• 1997

IgY (egg yolk immunoglobulin) against Helicobacter pylori was produced by immunizing hen with some Helicobacter pylori antigens. H. pylori whole cell, whole cell lysates, partially purified urease and p54 protein, which showed high antigenicity in mice, were used as immunogens. Four hens were immunized with these immunogens three times. IgY was purified from immunized egg yolk with polyethylene glycol (M.W. 8000) and its anti-H. pylori titer was determined by enzyme linked immunosorbent assay (ELISA). The anti-H. pylori titer reached peak at 8 weeks and was maintained over 20 weeks. H. pylori cells were agglutinated with these purified IgY and the specificity of these purified IgY was detected with immunoblotting.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.74-74
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• 2003

Embryonic stem cells have several characteristics suitable for cell replacement therapy. To investigate a possibility of using human embryonic stem cell (hESC) as a carrier of therapeutic gene(s), hESC (MB03) was co-transfected with cDNAS coding for tyrosine hydroxylase (TH) and GTP cyclohydrolase Ⅰ (GTPCH Ⅰ) and bulk-selected using neomycin and hygromycin-B. Successful transfection was confirmed by western immunoblotting and RT-PCR. The genetically modified hESC (bk-THGC) relieved apomorphine-induced asymmetric motor behavior by approximately 54% when grafted into striatum of 6-OHDA-denervated rat brain. The number of rotation, however, increased up to 176+18% in 6 weeks when sham-grafted compared with number of rotation before graft. Immunohistochemical staining revealed that the grafted hESC survived and expressed TH for at least 6 weeks while the experiment was continued.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.170-173
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• 2009

Vitellogenin (Vtg) is found in the serum of both female and male fish that have been exposed to environmental endocrine disrupters or estrogen hormones, and is used as a biomarker for such contamination. In our current study antibodies raised against the purified flatfish Vtg were tested for their reactivity on immunoblots to flatfish and carp Vtg, and also to a Vtg protein fragment produced in E. coli. Polyclonal antibodies raised against purified flatfish Vtg reacted well with Vtg in the serum of flatfish and carp induced with $17{\beta}$-estradiol, but not with the Vtg protein fragment produced in E. coli.

• Parasites, Hosts and Diseases
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• v.41 no.2
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• pp.121-123
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• 2003

This study was designed to detect and evaluate an antigenicity of low molecular weight proteins of Fasciola hepatica in fascioliasis. Low molecular weight protein of F. hepatica was purified by ammonium sulfate precipitation and Sephacryl S-100 HR gel filtration. The protein obtained was estimated to be 8 kDa on 7.5-15% gradient sodium dodecyl sulfate gel electrophoresis. Immunoblotting studies showed that the 8 kDa protein reacted with human fascioliasis sera, but not other trematodiasis sera. This result suggests that the 8 kDa protein of F. hepatica is one of diagnostic antigens in human fascioliasis without cross-reaction with other human trematodiasis.