• Title/Summary/Keyword: DNA-DNA hybridization

Search Result 871, Processing Time 0.026 seconds

Identification of Luteovirus Nucleotide Sequences in Mild Yellow-Edge Diseased Strawberry Plants

  • Shaban Montasser, Magdy;Al-Awadhi, Husain;Hadidi, Ahmed
    • The Plant Pathology Journal
    • /
    • v.18 no.1
    • /
    • pp.1-5
    • /
    • 2002
  • The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

Genomic Organization of ancop Gene for ${\alpha}-COP$ Homolog from Aspergillus nidulans

  • Lee, Hwan-Hee;Chae, Shun-Kee;Kim, Jeong-Yoon;Maeng, Pil-Jae;Park, Hee-Moon
    • Mycobiology
    • /
    • v.28 no.4
    • /
    • pp.171-176
    • /
    • 2000
  • We have cloned a ${\alpha}-COP$ homolog, ancop, from Aspergillus nidulans by colony hybridization of chromosome specific library using ${\alpha}-COP$ homologous fragment as a probe. The probe DNA was amplified with degenerated primers designed by comparison of conserved region of the amino acid sequences of Saccharomyces cerevisiae ${\alpha}-COP$, Homo sapiens HEP-COP, and Drosophila melanogaster ${\alpha}-COP$. Full length cDNA clone was also amplified by RT-PCR. Comparison of genomic DNA sequence with cDNA sequence obtained by RT-PCR revealed 7 introns. Amino acid sequence similarity search of the anCop with other ${\alpha}-COPs$ gave an overall identity of 52% with S. cerevisiae, 47% with human and bovine, 45% with Drosophila and Arabidopsis. In upstream region from the transcription start site, a putative TATA and CAAT motif were also identified.

  • PDF

Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
    • /
    • v.16 no.2
    • /
    • pp.77-82
    • /
    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

  • PDF

A Study on the Development of Electrochemical Biochip (전기화학적 바이오칩의 개발에 관한 연구)

  • Choi, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • Proceedings of the KIEE Conference
    • /
    • 2003.10a
    • /
    • pp.300-302
    • /
    • 2003
  • This research aims to develop the multi-channel type label-free DNA chip that has the above characteristics and be able to solve the problems. At first, we fabricated a high integrated type DNA chip array by lithography technology. It is able to detect a various genes electrochemically after immobilization of a various probe DNA and hybridization of label-free target DNA on the electrode s simultaneously.

  • PDF

Amino Acid Sequence Homology of Hybrid Poplar O-methyltransferuse Involved in Lignin Biosynthesis

  • Park, Young-Goo;Sul, Ill-Whan;Shin, Dong-Ill;Park, Jang-Won;Park, Hee-Sung
    • Journal of Plant Biotechnology
    • /
    • v.3 no.3
    • /
    • pp.131-134
    • /
    • 2001
  • In $\lambda$-Zap II vector system, a cDNA library was constructed for the developing secondary xylem mRNA from hybrid poplar, Populus nigra x maximowiczii. A cDNA clone of 1.5 kb in size, pOMTB1.4 encoding a lignin-bispecific O-methyltransferase was screened by plaque hybridization using a probe of 540 bp cDNA amplified by polymerase chain reaction from the cDNA library and identified by nucleotide sequencing. Its nucleotide sequence contains one open reading frame of 366 amino acids. The deduced amino acid sequence in comparison with that of Populus tremuloides showed the differences of 9 amino acids and revealed 85-99% homology among alfalfa, poplar and aspen.

  • PDF

Cloning and Characterization of the Putative Transferrin Receptor cDNA from the Olive Flounder (Paralichthys olivaceus)

  • Won Kyoung-Mi;Park Soo-Il
    • Fisheries and Aquatic Sciences
    • /
    • v.6 no.2
    • /
    • pp.101-104
    • /
    • 2003
  • A cDNA clone for the olive flounder (Paralichthys olivaceus) transferrin receptor (fTfR) was isolated from a leukocytes cDNA library. The fTfR gene consisted of 2,319 bp encoding 773 amino acid residues. The amino acid sequence alignment of the fTfR showed that their size and hydrophobic profile are similar. In addition, the Tyr-Thr-Arg-Phe (YTRF) motif that is the recognition signal for high-efficiency endocytosis, is conserved very well. This motif is important for functional properties of TfR. The deduced amino acid sequence had $42.4-42.9\%$ identities with the previously reported TfRs of vertebrates. The fTfR was expressed in the blood, kidney, spleen, and liver of healthy olive flounder by the Northern blot hybridization.

A Macrolide-Lincosamide-Streptogramin B Resistance Determinant Gene (ermJ) Cloned from B, anthracis 590

  • Kim, Hee-Sun;Choi, Eung-Chil;Kim, Byong-Kak;Park, Young-In
    • Archives of Pharmacal Research
    • /
    • v.15 no.1
    • /
    • pp.58-61
    • /
    • 1992
  • Bacillus anthracis 590 having an inducibla resistance determinant to MLS antibiotics was isolated from a soli sample in Korea. The resistance gene (ernJ) was cloned by Southern blotting of chromosomal DNA fragment digested by various restriction enzymes and coloy hybridization method and the cloned plasmid was named as pBA423. The size of inserted DNA fragment of pBS42 vector was about 2.9 kb and the DNA sequence of the subcloned fragment (Hinc II-Hinc II, 1.4kb) WAS determined. The DNA sequence of ernJ was composed of 357 bp for leader region and 861 bp for the structural gene. Because the leader sequence of ernJ was homologous to that of ermK, the expression of ernJ is also thought to be controlled by a transcriptionl attenuation mechanism.

  • PDF

Transgenic Tobacco Plants Introduced with cDNA of Cucumber Mosaic Virus Satellite RNA (오이 모자이크 바이러스 위성RNA의 cDNA가 도입된 형질전환 담배의 육성)

  • 이상용;홍은주;최장경
    • Korean Journal Plant Pathology
    • /
    • v.11 no.1
    • /
    • pp.80-86
    • /
    • 1995
  • The cDNA of CMV-As satellite RNA was introduced into tobacco plants (Nicotiana tabacum cv. Samsun NN) using a binary Ti plasmid vector system of Agrobacterium tumefaciens. The cDNA of satellite RNA introduced into tobacco plants was detected by polymerase chain reaction (PCR) and molecular hybridization analyses. Symptom development was distinctly suppressed in the transgenic tobacco plants when inoculated with CMV-Co. CMV concentration in the transgenic tobacco plants was decreased to 1/40 of non-transgenic tobacco plants. The kanamycin resistance gene of the transgenic tobacco plants was also detected in the progeny.

  • PDF

Cloning of cDNA Encoding the Precursor to the Small Subunit of Ribulose-1,5-Bisphosphate Carboxylase in Pea 9Pisum sativum) (완두콩(Pisum sativum)에서 Ribulose-1,5-Bisphosphate Carboxylase Small Subunit 유전자의 cDNA 클로닝과 광유도성 발현에 관한 연구)

  • 김한집
    • Journal of Plant Biology
    • /
    • v.32 no.1
    • /
    • pp.33-40
    • /
    • 1989
  • Polysomal polyadenylated mRNAs which were purified from pea leaves were fractionated by sucrose grandient sedimentation. Fractions corresponding to the peak at 11.5S were found to contain mostly mRNA encoding the precursor polypeptide to the small subunit of ribulose bisphosphate carboxylase (rbcS) by in vitro translation in wheat germ extract. Double-stranded cDNA which was synthesized from the 11.5S mRNA was cloned into Hind III site of plasmid pBR 325. A cDNA clone, H24, was identified to code for rbcS. In vitro translation product of the hybridization-selected mRNA was molecular weight 20,000, presumably the precursor of rbcS. The nucleotide sequences of the H24 showed almost complete homology with the sequences encoding the transit peptide of the rbcS-3A gene which was reported by Fluhr et al.(1986).

  • PDF

Tobacco plant transformed with a coat protein gene sequence of TMV (TMV외피 단백질 유전자의 연초로의 형질전환)

  • 이기원;박성원;김남원;박은경
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.15 no.2
    • /
    • pp.161-166
    • /
    • 1993
  • A double - stranded cDNA fragment (436bp) encoding coat protein of tobacco mosaic virus(TMV) was derived from the total 480nucleotides gene after reverse transcription of TMV RNA, and subclorled into a plant expression vector pBl 121, resulting in pBL 430. The plasmid DNA containing this chimeric gene was moved from E. cofi to Agrobacterium tumefaciens strain A28l, and was introduced in시 the tobacco plant by the Agrobocterium Ti - mediated transformtion system. The transformants were selected on a selection media containing kanamycin. The shoots add roots could be differentiated from the explants and whole plants were obtained. From Southern blot hybridization analysis, DNA extracted from transformants, it could be conformed that the chimeric gene fragment was inserted into the genomic DNA of tobacco plant.

  • PDF