• YAKHAK HOEJI
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• v.34 no.1
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• pp.11-14
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• 1990

$NH_2-terminal$ amino acid sequence analysis of monoclonal antibody is very important to identify gene family and diversities of antigen-antibody recognition. When we used the PVDF (Polyvinylidene difluoride) membrane blotting method, we could easily analyze $NH_2-terminal$ sequence of monoclonal-antibody which specifically binds to phosphatidylinositol 4,5-biphosphate. PVDF membrane is an ideal solid-phase support for sequence analysis, especially when used with electroblotting method. This method is superior to continual method and will be applied to the sequence analysis of picomole quantities of proteins by gel electrophoresis.

• Journal of the Korean Chemical Society
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• v.47 no.4
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• pp.363-369
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• 2003

Selenized yeast (Se yeast) containing $0.1{\%}$(w/w) of selenium was obtained when the yeast was incubated at a selenium concentration of 1$1.14{\times}10_-3 M$ in rich medium. After washing several times, the inorganic selenium on the cell wall was confirmed with MBRT. There was no indication of inorganic selenium on the cell wall when the blue color in MBRT was stayed for 15 minutes. The selenized yeast was sonicated, then the selenium contained protein was obtained after salting out by ammonium sulfate at the concentration $80{\%}$ saturation. The seven protein bands were seperated by SDS-PAGE and the selenium concentration in protein was measured by ICP-AES. Analytical data showed that the large expressed protein band contained a relatively large amount of selenium. The proteins of the 47kDa was contained the concentrations of 69.5 ${\mu}$ Se/g of most many content. The protein (47 kDa) was seperated from PVDF membrane by tank-electroblotting. The isolated protein was hydrolyzed under acid condition and reacted with PITC. The derivatives of amino acids were analyzed by HPLC and compared with the data obtained from regular yeast. The resulting selenium-yeast was analyzed with the selenomethionine concentration of $2{\%}$ comparaed with general amino acids. The goal of this study is to analyze the selenium concentration in protein bands and measure the degree of biotransformation of selenomethionine in a specific protein.

• The Korean Journal of Pesticide Science
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• v.10 no.1
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• pp.56-65
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• 2006

An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

• Parasites, Hosts and Diseases
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• v.31 no.1
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• pp.37-42
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• 1993

Surface proteins of Trichomonqs unginnlis (T vqsinalis) were analyzed to study the antigenic variation. The surface proteins of protozoa were labelled by N- hydrokysuccinimide-biotin (NHS-biotins, the NHS-biotin-labelled proteins were immunoprecipitated with rabbit antiserum to purifjr the antigenic fractions and analysed by SDS-PAGE plus electroblotting. The results obtained in this study were as follows; Biotinylated T. uaginalis-proteins obtained from intact cell and cells disrupted prior to labelling were detected by antibiotin-peroxidase in Western blots. Labelled proteins were immunoprecipitated by T. vcqinalis-immunized rabbit serum and the six bands with, the molecular weights of 46, 60, 68, 90, 130 and 220 kDa were identified as having antigenicity. T unginalis HY-1,HY-15 and ATCC 50148 were immunoprecipitated by immune rabbit serum after biotinylation and there were no difference from antigenic bands among these strains by this tehchnique. In conclusion with the results obtained in the present study, it was assumed that surface proteins of T vaqinclis were labelled by biotinylation and the six labelled bands at 46, 60, 68, 90, 130 and 220 kDa in their molecular weight were identified as having antigenicity by immunoprecipitation (IPI and this biotinylation-IP technique may be used for further study of surface antigen of T vaginalis.