• Title/Summary/Keyword: Probe DNA

Search Result 590, Processing Time 0.025 seconds

Electrophoretic Karyotyping by PFGE in the Genus Fusarium (Fusarium속에서 PFGE를 이용한 Electrophoretic Karyotyping)

  • Min, Byung-Re;Jung, Jin-Sook;Choi, Yong-Keel
    • The Korean Journal of Mycology
    • /
    • v.26 no.2 s.85
    • /
    • pp.135-143
    • /
    • 1998
  • Contour-clamped homogeneous electric field gel electrophoresis was used to establish electrophoretic karyotype for 10 species of Fusarium sections Sporotrichiella, Liseola, Gibbosum, Discolor and Martiella. Intact chromosomal DNA was isolated from fungal protoplast and separated under various conditions according to their size in order to improve DNA separation. The numbers of chromosome-sized DNA molecules for individual species ranged from 5-13, with individual chromosomes ranging from 0.78 Mb to 7.20 Mb in size. The total genome DNA size of each species was estimated at about 18.32 Mb to 48.20 Mb. Comparison of karyotype profiles following Southern hybridization analysis with a randomly selected genomic probe of F. oxysporum formae speciales litii was carried out.

  • PDF

Attribution of PAH Degradation of Sphingomonas chungbukensis DJ77 to the Plasmid pSY1 (Sphingomonas chungbukensis DJ77에 존재하는 Plasmid pSY1의 PAH 분해능)

  • 박승기;김성재;신희정;김영창
    • Korean Journal of Microbiology
    • /
    • v.37 no.2
    • /
    • pp.120-123
    • /
    • 2001
  • Sphingomonas chungbukensis DJ77 is able to use phenanthrene and biphenyl as the sole carbon and energy source. Mitomycin C curing experiment suggested that polyaromatic hydrocarbon (PAH) utilization in strain DJ77 was plasmid-encoded. The plasmid cured strains were failed to grow on the minimal medium sprayed with biphenyl or phenanthrene. This was evident from southern hybridizations using a previously cloned DNA segment as a probe. There were positive signals in the palsmid DNA of the wild-type strain DJ77 and the absence of hybridizations with chromosomal DNA from the plasmid DNA of the wild-type strain DJ77 and the absence of hybridizations with chromosomal DNA from the palsmid-cured mutant strains.

  • PDF

Integrated Type DNA Chip Array and Gene Detection Using an Indicator-free DNA (집적형 DNA칩 어레이 및 비수식화 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
    • /
    • 2006.07c
    • /
    • pp.1322-1323
    • /
    • 2006
  • This research aims to develop the multiple channel electrochemical DNA chip that has the above characteristic and be able to solve the problems. At first, we fabricated a high integration type DNA chip array by lithography technology. It is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously.

  • PDF

Development of an SH-SAW Sensor for Detection of DNA (DNA 측정용 SH-SAW 센서 개발)

  • Hur Youngjune;Pak Yukeun Eugene;Roh Yongrae
    • The Journal of the Acoustical Society of Korea
    • /
    • v.24 no.3
    • /
    • pp.160-165
    • /
    • 2005
  • We have developed SH (shear horizontal) surface acoustic wave (SAW) sensors for detection of the immobilization and hybridization of DNA (deoxyribonucleic acid) on the gold coated delay line of transverse SAW devices. The experiments of DNA immobilization and hybridization were performed with 15-mer oligonucleotides (probe and complementary target DNA). The sensor consists of twin SAW delay line oscillators operating at 100 MHz fabricated on $36^{\circ}$ rotated Y-cut $LiTaO_3$ piezoelectric single crystals. The relative change in the frequency of the two oscillators was monitored to detect the hybridization between target DNA and immobilized probe DNA in pH 7.4 PBS (phosphate buffered saline) solution. The measurement results showed a good response of the sensor to the mass loading effects of the DNA immobilization and hybridization with the sensitivity up to $1.55{\cal}ng/{\cal}ml/Hz$.

Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence In Situ Hybridization Technique

  • Han, M.S.;Cho, E.J.;Ha, H.B.;Park, H.S.;Sohn, S.H.
    • Reproductive and Developmental Biology
    • /
    • v.28 no.2
    • /
    • pp.133-137
    • /
    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

Actinodura roseorufa에서 생산되는 UK-58,852로부터 PKS type I 에 관련된 생합성 유전자의 분리 및 분석

  • Kim, Ja-Yong;Lee, Ju-Ho;Kim, Dae-Hui;Kim, Dong-Hyeon;Song, Jae-Gyeong;Lee, Hui-Chan
    • 한국생물공학회:학술대회논문집
    • /
    • 2000.11a
    • /
    • pp.660-664
    • /
    • 2000
  • To clone genes related UK-58,852 production, genomic DNA of strain Actinodura roseorufa was used for the construction of genomic library using pOJ446 cosmid vector. The genomic library was screened rising dehydratase PCR product and eryA gene as a DNA hybridization probe. pHD54 was isolated, which contained an approximately 35kb of inserted DNA. BamHI, SmaI and sonicater fragments hybridized to eryA probe. All of pHD54 BgmHI, SmaI and sonicater fragments were subcloned into pGEM7 and some fragments which hybridized to eryA probe were sequenced. The nucleotide sequence was analysed using BLAST program. The sequence identities were observed in KS,AT, KR, ER and PKS loading domains. Also oxidoreductase showed similarity to rifamycin module10, and dTDP-D-glucose 4,6 dehydratase and TDP-D-glucose synthase involved in biosynthesis of sugar showed similarity to Streptomyces argillaceus.

  • PDF

Molecular Cloning of $\beta$-Galactosidase Gene from Neisseria lactamica 2118 into Escherichia coli MC 1061 (Neisseria lactamica 2118의 $\beta$-galactosidase 유전자의 대장균으로의 클로닝)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
    • /
    • v.5 no.1
    • /
    • pp.37-45
    • /
    • 1992
  • The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

  • PDF

The Specific Probes Confirming the Genomic DNA of Tricholoma matsutake in Korea (송이의 Genomic DNA에 특이적인 Probe)

  • Lee, Sang-Sun;Hong, Sung-Woon;Chung, Hung-Chae;Sung, Chang-Kun;Kim, Jae-Hun;Ka, Kang-Hyeon;Kim, Hyun-Joong
    • The Korean Journal of Mycology
    • /
    • v.27 no.1 s.88
    • /
    • pp.20-26
    • /
    • 1999
  • The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

  • PDF

Development of Human Papillomavirus DNA Array by Using Lateral Flow Membrane Assay (Lateral Flow Membrane를 이용한 인유두종 바이러스 DNA Array의 개발)

  • Kim, Ki-Whang;Lee, Hyung-Ku;Cho, Hong-Bum
    • Korean Journal of Microbiology
    • /
    • v.44 no.4
    • /
    • pp.346-351
    • /
    • 2008
  • This study develops DNA array which can detect specific sequence of human papilomavirus (HPV) by using lateral flow membrane assay which is usually used for point of care test including pregnant diagnosis. Principle of HPV DNA array is as follow; fixing DNA probe which is peculiar to HPV type 6, 11, 16, 18, 31, 45 on a surface of lateral flow membrane and inducing hybridization response between probe and HPV PCR products which is obtained by using biotin-labeled MY09/l1 primers. And then, we can see the result of DNA hybridization that streptavidin labelled colloidal gold is responded with hybrid biotin. Lateral flow membrane array developed in this study confirms major HPV type economically and conveniently compared with existing HPV DNA chip method.

Construction of Shuttle Promoter-probe and Expression Vectors for Escherichia coli and Bacillus subtilis, and Expression of B. thuringiensis subsp. kurstaki HD-73 Crystal Protein Gene in the Two Species

  • Park, Seung-Hwan;Koo, Bon-Tag;Shin, Byung-Sik;Kim, Jeong-Il
    • Journal of Microbiology and Biotechnology
    • /
    • v.1 no.1
    • /
    • pp.37-44
    • /
    • 1991
  • A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, and the cp gene expression in B. subtilis begins early in the vegetative phase. The cell extracts from both clones were toxic to Hyphantria cunea larvae.

  • PDF