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Enzyme - Linked Immunosorbent Assay ( ELISA ) 법을 이용한 우유단백질의 열변성도 측정에 관한 연구
이승배(S . B . Lee),허경택(K . T . Huh),김창규(C . K . Kim),김종배(J . B . Kim),한석현(S . H . Han) 한국축산학회 1992 한국축산학회지 Vol.34 No.5
An enzyme-linked immunosorbent assay(ELISA) was developed to determine the extent of β-lactoglobulin (β-Lg) denaturation in heat treated milk. Polyclonal antibody was produced from the rabbit immunized with the β-Lg as an immunogen and was purified by the protein A affinity column. The antibody titer was determined above 1.28 × 10^5 in indirect ELISA. From the standard curve of indirect competitive ELISA for β-Lg. the sensitivitiy was found 90ng/㎖. Coefficient variations of intra- & inter-assay were 4.5-12% and 5.7-9.5%, respectively. When the ELISA of β-Lg were plotted against dilution of whey protein and skim milk, dilution assay curves could be made up to 40,000 times dilution of sample. When standard β-Lg was heated at 100℃ for longer time(5min. 15min & 30min). antibody-antigen reaction was decreased and the extent of heat-denatured β-Lg was reflected the difference of interval in dilution assay curve. When the whey protein and skim milk were heated respectively for various time(5min. l5min & 30min) at 100℃ the patterns of dilution assay curve were obviously displaced as compared with those of the whey protein and skim milk unheated and according to heating times, also made difference between dilution assay curves. When the skim milk was heated at various temperatures (65℃, 75℃ & 85℃) for 30min, according to higher temperature the patterns of dilution assay curve were showed apparent discrepancy against those of unheated skim milk.
한석현(S . H . Han),임영선(Y . S . Lim),김창규(C . K . Kim),이승배(S . B . Lee),권명상(M . S . Kwon),김종배(J . B . Kim) 한국축산학회 1993 한국축산학회지 Vol.35 No.3
This study was carried out to investigate the cleavage patterns of DQα class II MHC gene and uPA (urokinase type plasminogen activator) gene by southern blot analysis using human cDNA probe for DQα and uPA. From Sml of blood samples, 30∼45 ㎍ of genomic DNAs were prepared. The Abs_(260)/Abs_(280) ratios of each sample were 1.98 ∼2.08. The hybridization of uPA probe to rat genomic DNA digested with EcoR I exhibited 3.3kb and 1.25kb fragment. The hybridization of uPA probe to rabbit and bovine I genomic DNA exhibited 4.2kb and 1.25kb EcoR I fragment. But the bovine 2 genomic DNA showed only 5.45 EcoR I fragment. Southern hybridization of human uPA probe to genomic DNA of various species such as rat, rabbit and bovine digested with EcoR I revealed differences in RFLP pattern among species. Hybridization of DQα class II gene of bovine major histrocompatibility complex(MHC) to genomic DNA digested with BamH I showed the 3 major bands (7kb, 4kb, 0.96kb) and 1 minor band(2.7kb). The RFLP pattecn of bovines DQα gene was identical within species.
한석현(S . H . Han),박성현(S . H . Park),이정렬(J . L . Lee),김인정(I . J . Kim),김창규(C . K . Kim),이승배(S . B . Lee),권명상(M . S . Kwon),김종배(J . B . Kim) 한국축산학회 1993 한국축산학회지 Vol.35 No.4
We investigated the restriction fragment length polymorphisms(RFLPs) of 31 Korean native cattles and l4 beef cattles of three breeds(each 4 charolais, 5 angus, 5 hereford) with lysozyme cDNA(pBL-1) probe and several restriction endonucleases and tried to find differences between Korean native cattles and other beef cattles. When genomic DNAs were digested with restriction endonucleases (EcoRI, BamHI, PstI) each enzyme revealed different restriction fragment length polymorphisms (RFLPs). EcoRI digestion showed individual differences while other restriction enzyme digestions did not showed significant differences. When the genomic DNA were digested with EcoRI, two types of RFLPs were identified. One was single major hand pattern(A type) of 0.9kb and the other was double major band type(B type) of 0.9kb and 0.8kb. The type frequency of Korean native castles was different from that of other beef castles. Thirty of 31 Korean native castles were A type and only I was B type but 9 of 14 beef castles were A type and 5 were B type. When digested with BamHl, the gcnomic IJNAs revealed 9.0kb, 4.0kb and 0.8kb bands and digested with Pvu II 6.0kb, 4.0kb, 3.0kb and 1.5kb bands were identified.