• Current Research on Agriculture and Life Sciences
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• v.29
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• pp.37-42
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• 2011

The plant-homologue of Bax Inhibitor, a gene described to suppress the cell death induced by Bax gene expression in yeast, was isolated from rice (Oryza sativa L.). Nucleic acid sequence and amino acid sequence were 741 bp and 247 bp, respectively. The amino acid sequence of the predicted protein was well conserved in plant (84 % in amino acids) and contained five membrane-spanning segments.

• Korean Journal of Animal Reproduction
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• v.12 no.1
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• pp.51-62
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• 1988

Spleen cells of mouse immunized with hCG were fused with myeloma cell (SP 2/0 Ag 14) to produce monoclonal antibody against hCG. Several clones of hybridoma secreting monoclonal antibody were established and antibodies were characterized in terms of titer, subisotyping and sensitivity in immunoassay. Several methods, for the purification of anti¬bodies, based on gel-filtration, DEAE-ion exchange chromatography and affinity chromatography. were applied and compared each other by the result of SDS-PAGE. Two-site immunochemiluminometric assay (ICMA) involving the use of an excess concentration of a specific monoclonal antibody passively adsorbed onto the walls of plastic tubes and a chemiluminescence labelled antibody conjugate were de¬veloped for the determination of hCG as a preliminary study.

• Journal of Life Science
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• v.18 no.3
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• pp.298-303
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• 2008

Using a phage peptide library approach, we have isolated a peptide ligand that binds to transferrin receptor on the surface of human melanoma cell, B16F10. The library was first screened twice by recovering internalized phages and was further screened three times by competitively eluting transferrin receptor-specific phages with human transferrin among the phages bound to the cell surface. The peptides displayed by the selected phages were fused to translocation and catalytic domain of Pseudomonas exotoxin to prepare recombinant toxins. After estimating cytotoxicity of each recombinant toxin toward B16F10 cell, seven clones were selected. Sequence analysis revealed that one of the clones displayed a peptide which had a significant sequence homology with human transferrin. The peptide was chemically synthesized and was shown to be functional in delivering cytotoxic agents into B16F10 cell via interaction with transferrin receptor.

• Development and Reproduction
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• v.4 no.1
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• pp.67-77
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• 2000

Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

• Biomedical Science Letters
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• v.6 no.1
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• pp.73-82
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• 2000

Blunt-end DNA fragments can be inserted in two different orientations. Conventionally, their directions are determined by restriction enzyme digestion or by DNA sequencing, however, these methods are often limited in their use due to the lack of appropriate enzyme sites or large sample numbers, respectively. In the present study, a novel strategy and the corresponding protocol for the simple determination of insert orientation is introduced. Using conventional sequencing primers and PCR primers that have been used for amplification of the insert, single clones, which have inserted the fragment in the desired orientation, were easily identified by this PCR-based method. The fidelity of this system was confirmed by cloning of a tar urocortin cDNA, which is a recently discovered neuropeptide. Recombinant clones identified by this method were further shown to be fully functional, and using these, for the first time, urocortin was recombinantly expressed in eukaryotic cells.

• Polymer(Korea)
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• v.33 no.6
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• pp.602-607
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• 2009

A polyurethane (PU) surface enabling in vivo endothelialization via endothelial progenitor cell (EPC) capture was prepared for cardiovascular applications. To introduce CD34 monoclonal antibody (mAb) inducing EPC adhesion onto a surface, poly (poly (ethylene glycol) acrylate-co-butyl methacrylate) and poly (PEGA-co-BMA) were synthesized and then coated on a surface of PU, followed by immobilizing CD34 mAb. $^1H$-NMR analysis demonstrated that poly(PEGA-co-BMA) copolymers with a desired composition were synthesized. Poly(PEGA-co-BMA)-coated PU was much more effective for the immobilization of CD34 mAb, comparing with PEG-grafted PU prepared in our previous study, as demonstrated by that surface density and activity of CD34 mAb increased over 32 times. Physico-chemical properties of modified PU surfaces were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle, and atomic force microscopy (AFM). The results demonstrated that the poly(PEGA-co-BMA) coating was effective for CD34 mAb immobilization and feasible for applying to cardiovascular biomaterials.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.268-275
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• 2007

Diversity of the mud flat microbial population in Suncheon Bay was investigated by studying extracellular enzyme activities and 16S rDNA sequences. Four culturable bacterial strains with CMCase, xylanase and protease activities were isolated from the wetland and the mud flat. All the strains produced more xylanase activity than CMCase or protease activity, and the properties of the isolate enzymes from the wetland were similar to those from the mud flat. About 2,000 clones were obtained with the 16S rDNA amplified from the metagenomic DNA isolated from the mud samples. Based on the restriction pattern(s), seventeen clones were selected for base sequence analysis. Of the 17 clones, only 35% (6 clones) were found to be cultured strains and 65% (11 clones) to be uncultured strains. The similarities in the base sequences of the clones ranged from 91.0% to 99.9% with an average similarity of 97.3%. The clones could be divided into 7 groups, Proteobacteria (9 clones, 52.9%), Firmicutes (3 clones, 17.6%), Bacteroidetes (1 clone), Flavobacteria (1 clone), Verrucomicrobia (1 clone), Acidobacteria (1 clone), and Chloroflexi (1 clone). Most of the Proteobacteria clones were gamma Proteobacteria associated with oxidation-reduction of sulfur.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.489-493
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• 1989

The role of each supplement in serum-free medium KM3 for the growth of hybridoma and the production of monoclonal antibody was investigated. Transferrin, ethanolamine and bovine serum albumin were shown to be indispensable for the growth of four kinds of hybridoma tested in this work, especially transferrin for Alps 25-3, and ethanolamine for A4W and KW hybridoma. The addition of $\beta$-mercaptoethanol to the culture medium of HCGK showed a good influence of both the cell growth and the production of monoclonal antibody. Upon the experimental results, we suggested a serum-free medium containing a minimum composition for the culture of hybridoma.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.371-384
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• 2007

연구 목적 : 면역 반응에서 B-1 세포의 정확한 역할은 아직 명확히 규명되지 않았으나 최근 B-1 세포가 면역의 내성을 야기하고 유지하는데 필요한 특성들을 가지고 있음이 밝혀지고 있다. 이에 본 연구에서는 B-1 세포 유래 단클론 항체의 특성과 항원 자극에 대한 B 세포의 동시자극자 (MHC Class, B7-2, 7-2) 발현을 평가하여 B 세포의 면역조절 기능을 알아보고자 한다. 연구 대상 및 방법 : B-1 세포 유래 단클론 항체를 형성하는 잡종세포주를 이용하여 다양한 내, 외인성 항원에 대한 단클론 항체의 반응 양상을 평가하였다. 여러 내, 외인성 항원으로 면역한 쥐의 복강과 비장 B 세포의 동시자극자의 발현을 Fluorescence Activated Cell Sorting (FACS)을 이용하여 평가하였다. 결과 : 최종적으로 단클론을 형성하는 2개의 클론을 형성하였고, 이 B-1 세포 유래 단클론 항체는 dose-saturable pattern을 띄는 다중 반응성을 나타내었다. FACS를 이용한 동시자극자의 발현 검사에서는 MHC 발현은 복강과 비장의 B 세포가 유사하였으나, B7-1과 7-2는 복강의 B 세포에서 더 뚜렷한 발현을 보여주었다. 결론 : B-1 세포 유래 단클론 항체는 다양한 내, 외인성 항원 자극에 대해 dose-saturable한 다중반응성을 나타낸다. 복강과 비장의 B세포는 내, 외인성 항원의 면역에 있어서 동시자극자 발현이 명확히 다른 양상을 나타냄을 확인할 수 있었다.

• Food Science of Animal Resources
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• v.28 no.5
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• pp.651-659
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• 2008

This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.