• Title/Summary/Keyword: hybridization

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Comparative analyses of Theileria sergenti isolated from Korea and Japan by southern hybridization and polymerase chain reaction (Sourthern hybridization과 중합효소연쇄반응을 이용한 한국과 일본의 Theileria sergenti 비교)

  • Chae, Joon-seok;Lee, Joo-mook;Kwon, Oh-deog;Lee, Seung-ok;Chae, Keon-sang;Onuma, Misao
    • Korean Journal of Veterinary Research
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    • v.36 no.1
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    • pp.187-193
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    • 1996
  • The T sergenti DNA fragments used as probes of KTS1(2.4kb) and KTS3(1.5kb) were labeled with digoxigenin-11-dUTP for the Southern hybridization. T sergenti DNAs from different geographic locations(Korea; Chonbuk, Kyungbuk, Chungnam, Kangwon, Cheju island, Japan; Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9202, Shintoku 9102) which had been digested with Pst I and EcoR I were probed by the digoxigenin-11-dUTP-labeled KTS1 and KTS3. As the results, the samples from Chonbuk, Kyungbuk, Cheju island in Korea and Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9102 in Japan were positively reacted, but the others from the other locations not reacted. In the comformation test of T sergenti DNA from different geographic locations, all of the samples were positively detected by PCR amplification.

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Identification of Aujeszky's disease virus by in situ hybridization (In situ hybridization 조직화학법을 이용한 오제스키병 바이러스 동정)

  • Kim, Soon-bok;Sur, Jung-hang;Moon, Oun-kyung
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.327-333
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    • 1994
  • The purpose of this study was to establish a rapid diagnostic method detecting Aujeszky's disease virus (ADV) DNA in the cultured cell monolayers (PK-15) and tissue sections of ADV(NYJ-1-87)-infected rats and pigs by in situ hybridization(ISH). Detection of specific ADV-DNA in infected cells was conducted by radiolabeled ISH method using $^{32}P-labeled $ DNA probe (BamH1 7 fragment) which contains a 6.3 Kb ADV-DNA insert. Where ADV-DNA was detected by radiolabeled ISH, the deposition of black photographic grains occurred in the nuclei and the cytoplasms of ADV-infected cells. Positive hybridization signal was often observed in the spinal trigerminal nucleus of the pons, the nucleus of the trigerminal ganglion neuron and the epithelial cells of tonsillar crypts. The results suggested that ISH is considered as a highly sensitive and reliable tool for confirmative diagnosis of this viral disease.

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Neuroanatomical Localization of Cells Containing Gonadotropin Releasing Hormone mRNA in the Brain of Frog, Rana dvbowskii, by in situ Hybridization (In situ hybridization법에 의한 북방산개구리 뇌에서 GnRH mRNA를 함유한 세포의 분포 연구)

  • 최완성;김정우
    • The Korean Journal of Zoology
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    • v.37 no.3
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    • pp.304-310
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    • 1994
  • Using in situ hybridization, we have mapped the anatomical localization of perikarya containing myNA that codes for sonadotropin releasing hormone (GnRH) in the brains of female frogs, R. dybowskii. DNA olisomers, with sequences complementary to the GnRH portion of pro-GnRH myNA sequence, were synthesized and hybridized to paraformaldehvde-fixed, sagittal sections of the whole brain stem. The distribution of the GnRH mRNA containing cell bodies was similar to that described for GnRH peptide by immunohistochemistrv. That is, cells containing GnRH mRNA were observed in the medial septal area, anterior preoptic area, ventromedial hvpothalamus and infundibular regions. However, another cell groups which contains GnRH mRNAs were also detected by in situ hybridization in the bed nucleus of hippocampal commissure, preoptic area, nucleus infundibularis dorsalis, mesencephalic nuclei and intermediolateral cell column of spinal cord areas. These results demonstrate the feasibility of using in situ hybridization as a strategy to study the distribution of GnRH neurons and the detection of GnRH gene expression in the vertebrates.

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Cloning of nif genes from Enterobacter agglomerans in Escherichia coli. (Enterobacter agglomerans의 질소고정유전자 Cloning)

  • 정건섭;이정기;민태익;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.116-121
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    • 1987
  • In order to cloning of the nif genes of Enterobacter agglomerans NFB-264, the digested total DNA of the strain was ligated to pBR 322 and transformed into E. coli. Through the negative selection and colony hybridization, the transformants were obtained. The recombinant plasmids, pNEL 10 and pNES 20 were extracted from these transformants. It was known from Southern hybridization that pNEL 10 contained the 12 Mdal foreign DNA fragment hybridized with nif Q-X probe and pNES 20 included the 5 Mdal foreign DNA fragment hybridized with nif NE and nif YK probe.

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DNA Hybridization Simulation with Single Base Mismatches for DNA Computing (1-Base non Watson-Crick 결합을 허용하는 DNA Hybridization Simulation)

  • 장하영;신수용;장병탁
    • Proceedings of the Korean Information Science Society Conference
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    • 2003.04c
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    • pp.476-478
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    • 2003
  • 1-Base의 non Watson-Crick 결합과, Dangling end(결합이 이루어진 두 개의 DNA strand 중 한쪽 끝이 다른 쪽 끝보다 짧은 경우)를 허용하는 nearest-neighbor model을 사용하여 DNA/DNA Hybridization 예측 시스템을 구현하였다. DNA 컴퓨팅을 기존의 실리콘 컴퓨터를 이용하여 접근하는 이러한 방법은 좀 더 효율적인 분자 알고리즘의 개발과 DNA 컴퓨팅에 사용될 수 있는 더욱 신뢰성 있는 DNA 시퀀스의 설계에 도움을 줄 수 있을 것이다.

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HEMAGGLUTINATION AND COLONY HYBRIDIZATION FOR THE IDENTIFICATION OF ENTEROTOXIGENIC Escherichia coli ISOLATED FROM HEALTHY PIG

  • Choi, S.H.;Oh, M.J.;Sung, C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.671-677
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    • 1996
  • Erythrocytes from three different animal species were used to determine mannose-sensitive hemagglutination (MSHA) and mannose-resistant hemagglutination (MRHA) of 755 isolates obtained from rectal swabs of healthy pig. In addition, colony hybridization using digoxigenin-dUTP labeled polynucleotide probes was performed for the detection of heat-stable and heat-labile enterotoxin genes carried by MRHA positive isolates. Of 755 strains, 9, 4 and 28 strains gave a positive MRHA with bovine, equine and pig erythrocytes, respectively. Of these isolates, 28 (3.7%) were characterized for positive MRHA by at least one blood. Seven isolates gave a positive MRHA with two kinds of blood. Three gave a positive MRHA with three kinds of blood. Twenty-eight strains, while positive in MRHA, yielded negative signals in the colony hybridization assay for the detection of heat-stable (STaI and STaII) and heat-labile (LT) enterotoxin genes in E. coli.

Detection of cryIB Genes in Bacillus thuringiensis subsp. entomocidus and subsp. subtoxicus

  • CHOI, SOO KEUN;BYUNG SIK SHIN;BON TAG KOO;SEUNG HWAN PARK;AND JEONG IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.171-175
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    • 1994
  • To find new crystal protein genes, we screened 42 Bacillus thuringiensis strains of serovar standards by Southern hybridization with a cryI-specific probe which was amplified from B. thuringiensis subsp. kurstaki HDl by polymerase chain reaction (PCR). Two strains, B. thuringiensis subsp. entomocidus HD9 and subsp. subtoxicus HD109, generated weak signals under the low-stringency hybridization conditions. Further analysis with Southern hybridization revealed that the two strains contained cryIB genes which are slightly different from those of B. thuringiensis subsp. thuringiensis HD2. These results were confirmed by PCR with cryIB-specific primers followed by the restriction analysis of PCR products.

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Enumeration of Vibrio vulnificus in Natural Samples by Colony Blot Hybridization

  • Lee, Jeong-Hyun;Lee, Kyu-Ho;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.302-309
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    • 2001
  • Colony blot hybridization using a VVHP DNA probe derived from the sequence of the hemolysin gene, vvhA, was specific in identifying all V. vulnificus strains, thereby, eliminating the need for any additional phenotypic identification. The colony blot hybridization procedure revealed a sensitivity and broad applicability sufficient for the direct enumeration of V. vulnificus in various natural samples, without the use of enrichment or culturing on selective medis. V. vulnificus was detected in all natural samples collected during August and May at concentrations ranging from $2.1{\times}10^1\;to\;4.0{\times}10^3$ organisms per ml. However, during November and February, when the mean temperatures of the seawater were $12^{\circ}C$ and $5^{\circ}C$, respectively, V. vulnificus was not detected in any natural samples.

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Hybridization에 의한 반수체 재조합 효모균주의 전분 발효능 증진

  • Park, Sun-Young;Kim, Keun;Lee, Chang-Hoo
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.726-732
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    • 1996
  • To improve the fermentation characteristics(such as starch-degradability, ethanol tolerance, sugar and high-temperature tolerance) of recombinant haploid yeast Saccharomyces diastaticus K114, hybridization technique was used. The hybridization partner was S. diastaticus 1177 which had good glucoamylase activity and fermentabi- lity. The best hybrid HH64 showed improved ethanol tolerance, sugar and high-temperature tolerance. Especia- lly, the starch-fermentability was significantly improved, since the hybrid produced 1.60% (w/v) ethanol from 4% (w/v) starch, while the recombinant haploid K114 produced 1.30% (w/v) ethanol. The optimum temperature and pH for the starch-fermentation by the hybrid HH64 was 30$\circ$C and 5, respectively. The hybrid yeast HH64 produced 7.5% (w/v) ethanol directly from 20% (w/v) starch.

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Development of dot blot hybridization method using non-radio labeled probes for the diagnosis of malignant catarrhal fever (Dot blot hybridization에 의한 malignant catarrhal fever virus의 진단법 개발)

  • Kim, Ok-Jin
    • Korean Journal of Veterinary Pathology
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    • v.7 no.1
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    • pp.1-4
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    • 2003
  • Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Dot blot hybridization (DBH) protocols for detecting and differentiating this MCF virus were developed. OvHV-2 specific primer pairs, 556/555, were used for the amplification of target DNA. Then, the amplified DNA was labeled with incorporation of digoxigenin (DIG). The Dig-labeled probe was able to detect and differentiate specifically OvHV-2 DNA. This DBH technique can be applied to confirm the presence of MCF virus on clinical samples and to differentiate specifically between OvHV-2 infection and other viral infections.

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