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염전위충(Haemonchus contortus) 생식기관의 DNase활성에 관한 연구
곽동미,Kwak, Dongmi 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3
DNase activity in Haemonchus contortus reproductive tissue was characterized and compared to that in whole worm. DNase activity in reproductive tissue was detected throughout pHs 4-10 with high activity under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. The activity produced DNA fragments with mixtures of 3'-hydroxyls (OH) and 3'- phosphates (P) at each pH. Three distinct DNase activities were identified and had $M_rs$ of 34, 36 and 38.5 kDa in zymograms, which were distinguished according to pH requirement and sensitivity to EDTA. Among them, the 36 kDa reproductive tissue DNase had predominant activity at pH 5.0, but very weak at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 36 kDa reproductive tissue DNase resemble those of classic acidic DNases. In contrast, 36 kDa whole worm DNase activity had high activity at both pH 5.0 and 7.0. While the 36 kDa DNase activity at pH 5.0 was similar in both reproductive tissue and whole worm samples, the activity at pH 7.0 was predominantly detected in whole worm sample. This suggests that the 36 kDa whole worm DNase at pH 5.0 differs from that at pH 7.0. Thus, results indicate that the EDTA-insensitive 36 kDa DNase at pH 5.0 is specific for H. contortus reproductive tissue.
Fenbendazole에 저항성과 감수성을 지닌 염전위충의 분비배설물에서의 DNase 활성 비료
곽동미,Kwak, Dongmi 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3
Change in ${\beta}$-tubulin nucleic acid and protein sequences was the only known difference between Haemonchus contortus fenbendazole (FBZ)-resistant and -susceptible isolates. This change was sufficient to determine the pathologic effect induced by FBZ treatment. This research was initiated to investigate further differences from these two isolates. Since ${\beta}$-tubulin is involved in formation of microtubule, which has functions in secretory vesicle transport, DNase activities from excretory/secretory products (ESP) of the two isolates were compared, based on pH, sensitivity to DNase inhibitors, molecular masses and production of 3'-OH. The most significant difference detected was that a 38.5 kDa DNase activity was identified from ESP of H. contortus FBZ-susceptible isolates but not from those of H. contortus FBZ-resistant isolates. However, it was shown that the 38.5 kDa DNase is expressed with similar level of activity in intestine and whole worm of H. contortus FBZ-resistant and -susceptible isolates. This result demonstrated that the secretory transport pathway of the 38.5 kDa DNase was inhibited by unknown mechanisms, which may be related with ${\beta}$-tubulin sequence change in FBZ-resistant isolates. Other DNases of 34, 36 and 37 kDa were detected from ESP of both H. contortus FBZ-resistant and -susceptible isolates. Overall DNase activities found from ESP of these two isolates were not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. In addition, DNase activities in two isolates produced DNA fragments with mixtures of 3'- hydroxyls (OH) and 3'-phosphates (P) at each pH although the 3'-end labeling ratios at pH 5.0 and 7.0 were shown different. Identification of inhibition of the 38.5 kDa DNase secretion in FBZ-resistant isolates suggests existence of further differences, in addition to ${\beta}$-tubulin sequence change, in two isolates. This shows complex effect of FBZ on H. contortus biological mechanisms.
곽동미,권오덕 한국임상수의학회 2004 한국임상수의학회지 Vol.21 No.3
This studies was carried out to investigate a process of formation for the granulomatous lesions in the liver and the haematological variation with the lapse of time after infection of Capillaria hepatica in dogs. Twelve crossbred puppies, about 3 months of age and 2-3 kg of body weight, were administered with 2,000 Capillaria hepatica infective eggs. Every four puppies was sacrificed on 1 week, 3 weeks and 5 weeks after infection, respectively. Although no marked clinical sign was noticed, total leukocyte values were increased peak on 1 week, and then reduced gradually on 3 weeks and 5 weeks after infection. Absolute differential counts of neutrophils and lymphocytes were significantly increased on 1, 3 and 5 weeks after infection. Absolute differential counts of monocytes and eosinophils were trend to increase during the experimental periods. On grossly findings, liver congestions were observed in all infected puppies, and a few white specks were scattered under liver capsule in one puppy on 3 weeks and two puppies on 5 weeks after infection. On microscopic findings, many fresh larvae were observed in the liver tissues in one puppy on 1 week after infection. A worm was decayed and only a portion of cuticle was shown in one puppy on 3 weeks after infection. Around the central necrotic material, the layers of thick macrophages with a few giant cells and lymphocytes with fibrous connective tissues were consisted the granulomatous lesions on 5 weeks after infection.
곽동미 한국임상수의학회 2004 한국임상수의학회지 Vol.21 No.3
DNase activity from infective larvae of the parasitic nematode Haemonchus contortus was characterized andcompared to that from whole worm. DNase activity from infective larvae was detected throughout pHs 4-10, but highactivity was detected under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but wassignificantly inhibited at pH 7.0. The activity produced DNA fragments with mixtures of 3'-hydroxyls (OH) and 3'-larvae on zymograms. The 37 kDa DNase was detected only at pH 5.0, but not at pH 7.0, and this activity was notinhibited by EDTA at pH 5.0. These characteristics of the 37 kDa infective larval DNase resemble those of classic acidicDNases (e.g., DNase II). In contrast, 34, 36 and 38.5 kDa DNase activities were shown to be specific for whole worm.This result demonstrated that DNases in H contortus are regulated during development.
Babesia equi ema-l 5' intergenic 뉴클레오타이드의 프로모터 위치 확인: I. PCR 증폭 및 제한효소지도
곽동미 한국동물위생학회 2004 韓國家畜衛生學會誌 Vol.27 No.1
Babesia equi ema-1 5' intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5' IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes (Bgll , HindⅢ, Kpn1 and BamH1) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.
Babesia euqi ema-1 5` intergenic 뉴클레오타이드의 프로모터 위치 확인: I. PCR 증폭 및 제한효소지도
곽동미 ( Dong Mi Kwak ) 한국동물위생학회 2004 한국동물위생학회지 (KOJVS) Vol.27 No.1
Babesia equi ema-1 5` intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5` IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes(Bgll, HindⅢ, Kpnl and BamHl) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.