• Title/Summary/Keyword: Fragment method

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Detection of Dirofilaria immitis by Reverse Transcription Polymerase Chain Reaction in Canine (역전사중합효소연쇄반응을 이용한 개심장사상충의 검출)

  • 이영준;박진호;권오덕;이주목
    • Journal of Veterinary Clinics
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    • v.16 no.1
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    • pp.177-181
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    • 1999
  • This study was undertaken to clarify the more accurate detecting method of Dirofilaria immitis. Seven dogs, average 7.47 years old, confirmed with Dirofilaria immitis infection by modified Knott's method were used as the experimental animals. cDNA was constructed using oligodT(15) primer after extracting total RNA from the blood of dogs that were confirmed with Dirofilaria immitis infection. As a result of polymerase chain reaction with template using constructed cDNA, the predicted products of a 378 base-pair DNA fragment was amplified. From these results, RT-PCR was more sensitive and effective than modified Knott's method to detect Dirofilaria immitis in dogs.

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Detection and Identification of Mycobacterium Tuberculosis in Patients with Tuberculous Cervical Lymphadenitis by PCR-RFLP (경부 결핵성 임파선염 환자에서 PCR-RELP를 이용한 결핵균의 검출 및 확인)

  • Lee Sang-Sook;Cho Young-Rok;Chun Ji-Min;Choi Yong-Seok;Sohn Eun-Ju;Park Nam-Cho;Park June-Sik
    • Korean Journal of Head & Neck Oncology
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    • v.12 no.2
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    • pp.169-176
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    • 1996
  • Tuberculous cervical lymphadenitis is still an important cause of neck mass in Korea. Tuberculosis is an important differential diagnosis in patients of cervical lymphadenopathy. Rapid and sensitive test for the diagnosis of tuberculosis is essential for the approapiate treatment. Up to now, conventional diagnostic methods for M. tuberculosis were acid-fast bacilli(AFB) stain and culture of M. tuberculosis. The direct microscopic examination of AFB by Ziehl-Neelsen stain is rapid, but often negative. The culture for M. tuberculosis is time-consuming, taking 4 to 8 weeks. Recently various methods to detect Mycobacterial DNA, including PCR and restriction fragment length polymorphism(RFLP) analysis have been reported. Here we represent a simple method for the confirmation of M. tuberculosis and exclusion of the other Mycobacterial species by RFLP analysis and silver staining of polyacrylamide gel electrophoresis after nested PCR for a repetitive DNA sequence(IS986) specific for M. tuberculosis from fresh or paraffin-embedded biopsy specimens. This result leads us to conclude that this method is simple, rapid and possibly applicable to confirm M. tuberculosis and rule out the other Mycobacteria species from the clinical specimens in the clinical laboratories.

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Cloning and Characterization of a Gene for Fibrinolytic Enzyme from Bacillus subtilis BB-1 Isolated from Black Bean Chung-kuk (흑두로 제조한 청국에서 분리된 Bacillus subtillus BB-1으로 부터 혈전용해효소 유전자 크로닝 및 특성규명)

  • Lee Young-Hoon;Lee Sung-Ho;Jeon Ju-Mi;Kim Hong-Chul;Cho Yong-Un;Park Ki-Hoon;Choi Young-Ju;Gal Sang-Wan
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.513-521
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    • 2005
  • A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.

Development of a Species Identification Method for the Egg and Fry of the Three Korean Bitterling Fishes (Pisces: Acheilognathinae) using RFLP (Restriction Fragment Length Polymorphism) Markers (제한절편 길이 다형성(RFLP) 분자마커를 이용한 납자루아과 담수어류 3종의 난과 치어 종 동정 기법 개발)

  • Choi, Hee-kyu;Lee, Hyuk Je
    • Korean Journal of Environmental Biology
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    • v.36 no.3
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    • pp.352-358
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    • 2018
  • This study aimed to develop a species identification method for the egg and fry of the three Korean bitterling fishes (Pisces: Acheilognathinae), including Acheilognathus signifer, Acheilognathus yamatsutae and Rhodeus uyekii based on the PCR-based Restriction Fragment Length Polymorphism (RFLP) markers. We conducted a field survey on the Deokchicheon River from the North Han River basin, where the three Acheilognathinae species co-occur, and also analyzed the existing sequence dataset available from the GenBank. We found coexistence of the three species at the study site. The egg and fry were obtained from the host mussels (Unio douglasiae sinuolatus) by hand from May to June 2015 and in May 2017. To develop PCR-based RFLP markers for species identification of the three Acheilognathinae fish species, restriction enzymes pinpointing species-specific single nucleotide variation (SNV) sites in mitochondrial DNA COI (cytochrome oxidase I) and cyt b (cytochrome b) genes were determined. Genomic DNA was extracted from the egg and fry and RFLP experiments were carried out using restriction enzymes Apal I, Stu I and EcoR V for A. signifer, A. yamatsutae and R. uyekii, respectively. Consequently, unambiguous discrimination of the three species was possible, as could be seen in DNA band patterns from gel electrophoresis. Our developed PCR-based RFLP markers will be useful for the determination of the three species for the young and would assist in studying the spawning patterns and reproductive ecology of Acheilognathinae fishes. Furthermore, we believe the obtained information will be of importance for future maintenance, management and conservation of these natural and endangered species.

Determination and confirmation of the carbendazim residue in soybean sprout (콩나물중 살균제 carbendazim 잔류분의 정량 및 확인)

  • Kim, Young-Gook;Park, Jong-Tae;Hong, Suk-Soon
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.79-84
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    • 1998
  • Tandem HPLC and atmospheric pressure chemical ionization(APcI) LC/MS method was used for the determination and confirmation of carbendazim residues in soybean sprout. Fluorescence(FL) detector was connected in tandem with the ultraviolet(UV) detector for dual detection of the carbendazim residue at the excitation and emission wavelength of 280 nm and 310 nm, respectively. The limit of detection for carbendazim was $0.1{\mu}g/kg$ sample. Recoveries of carbendazim from fortified soybean sprout at 0.5, 1.0 and 2.0 ppm were averaged 89.1%. Mass spectrometry using a APcI source confirmed the carbendazim residue in the soybean sprout sample. Fragmentation pattern on the APcI LC/MS spectrum of carbendazim was simpler than that from electron impact(EI) mass spectrum. Carbendazim produced 3 major ions including m/z 133, m/z 159 and m/z 191($M^{+}$). This method was sensitive enough to provide reliable and reproducible results for practical applications.

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Identification of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Golpayegani Cattle by DNA Test

  • Mosafer, J.;Nassiry, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1691-1695
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    • 2005
  • The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

Asynchronous and Adaptive Massage Passing Scheme for Wireless Sensor Networks (무선 센서 네트워크를 위한 비동기 적응형 메시지 전달 기법)

  • Jeong, Jong-Kyun;Ra, In-Ho
    • Journal of the Korean Institute of Intelligent Systems
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    • v.23 no.3
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    • pp.196-201
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    • 2013
  • In this paper, we propose an asynchronous and adaptive message passing scheme based on S-MAC for handling with the problem on energy efficiency in wireless sensor networks. The proposed scheme consists of a policy that differentiate transmission errors caused by packet collision or radio interference from congestion errors caused by buffer overflow and a method that adaptively controls the size of a fragment according to the variance of traffic loads. Especially, it presents a method that highly reduce the energy consumption by keeping the size of fragment not being excessively smaller than the one that may result in rapid increment of the total transfer time. Finally, with the simulation results we show that network throughput and delay are improved by using the proposed message passing scheme.

Analysis of genetic relationships of Colletotrichum spp. isolated from sweet persimon with AFLP (AFLP를 이용한 단감나무 탄저병 병원균 Colletotrichum spp.의 유연관계 분석)

  • Kim, Hee-Jong;Jeong, Bong-Gu;Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.9-14
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    • 2001
  • Colletotrichum species are important fungal pathogens that cause great damages on various host plant species worldwide. In Korea, Colletotrichum species cause massive economic losses on apple, peach, grape, and especially, sweet persimon productions. In the past, identification of the pathogen and the studies on the genetic relationships among the pathogenic isolates were mainly based on morphology, cultural characteristics, and the difference in pathogenicity. However, in recent years, these traditional methods have been replaced with molecular methods including AFLP. AFLP method with the merits of both RAPD and RFLP has been widely used for the genetic relationship studies of various organisms. Therefore, in this study, AFLP method was employed for the studies of genetic relationships among the different isolates of Colletotrichum species collected from various parts of sothern Korea. As a result, two specific band pattern groups were observed among different isolates of Colletotrichum species.

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The Expression of Fantastic Body in Fashion Illustration (패션 일러스트레이션에 나타난 판타스틱 신체의 표현 분석)

  • Choi, Jung-Hwa
    • Fashion & Textile Research Journal
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    • v.11 no.6
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    • pp.867-877
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    • 2009
  • These days, the fantastic in opposition to classic beauty becomes a genre of creative body expression. The purpose of this study was to analyze the expressive characteristics of body types and meanings in recent fantastic fashion illustration. The method of this study was to analyze recent documentaries, fashion books, internet web site and so forth. The results were as follows: In literatures, pictures and movies, the category of the fantastic body's expressive types were classified as dominant mutant based on SF, multi body or fragment body by disruption, heterogeneous compound based on myth, personified humanoid and non substance in supernatural boundary. The dominant mutant based on SF was expressed image morphing, composition of machine image with body and modern metamorphosis of classic SF body. It means propensity to post-feminism and reservation of meaning analysis based on human unconsciousness. The multi body or fragment body by disruption in fashion illustration was expressed distorted composition of same body pictures, replacement of different bodies, deconstruction and partial omission of body and composition of meaning or non meaning images. It means permanence of self and basic narcissism. The heterogeneous compound based on myth was expressed general composition or optical illusion of various and aggressive animal motive. It means reinterpretation of original myth, metaphor of basic femme fatale, pursuit of permanence and sign of primitive mind in unconsciousness. The personified humanoid was expressed real human body description of mannequin or ball joint doll and anthropomorphism of robot image. It means representative satisfaction and nostalgia of childhood. The non substance in supernatural boundary was expressed grotesque description of ghost, zombie, vampire, angel, fairy, using of symbolic red, black color and non body. It means human's basic desire about immortality and taboo. Through the result of these study, the expression of fantastic body in fashion illustration will expend expressive method and we will understand human and cultural codes of today.

Identification of triacylglycerols in coix seed extract by preparative thin layer chromatography and liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry

  • Sim, Hee-Jung;Lee, Seul gi;Park, Na-Hyun;Kim, Youna;Cho, Hyun-Woo;Hong, Jongki
    • Analytical Science and Technology
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    • v.30 no.2
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    • pp.102-111
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    • 2017
  • Here we reported a methodology for identification of triacylglycerols (TAGs) and diacylglycerols (DAGs) in coix seed by preparative thin layer chromatography (prep-TLC) and non-aqueous reversed-phase liquid chromatography (NARP LC)-atmospheric pressure chemical ionization (APCI) tandem mass spectrometry (MS/MS). Lipid components were extracted from coix seed by reflux extraction using n-hexane for 3 hr. TAGs and DAGs in coix seed extract were effectively purified and isolated from matrix interferences by prep-TLC and then analyzed by LC-APCI-MS and MS/MS for identification. TAGs were effectively identified taking into consideration of their LC retention behavior, APCI-MS spectra patterns, and MS/MS spectra of $[DAG]^+$ ions. In MS/MS spectra of TAGs, diacylglycerol-like fragment $[DAG]^+$ ions were useful to identify TAGs with isobaric fragment ions. Based on an established method, 27 TAGs and 8 DAGs were identified in coix seed extract. Among them, 15 TAGs and 8 DAGs were for the first time observed in coix seed. Interestingly, some of TAGs isolated by prep-TLC were partly converted into DAGs through probably photolysis process during storing in room temperature. Thus, degradation phenomenon of TAGs should be considered in the quality evaluation and nutritional property of coix seed. LC-APCI-MS/MS combined with prep-TLC will be practical method for precise TAG and DAG analysis of other herbal plants.