• Title/Summary/Keyword: DNA-DNA hybridization

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RFLP(Restriction Fragment Length Polymorphism) by Ribosomal RNA and M13 Probes of Clostridum thermocellum Strains (Ribosomal RNA와 M13 probe에 의한 clostridium thermocellum 균주들의 RFLP(Restriction Fragment Length Polymorphism)비교)

  • 이호섭;홍수형;하지홍
    • Korean Journal of Microbiology
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    • v.29 no.3
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    • pp.189-194
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    • 1991
  • The degree of the genetic variations among Clostridium thermocellum ATCC 27405 and the wild type strains was investigated by the mehtod of GC ratio, DNA-DNA hybridization and RFLP (Restriction Fragment Length Polymorphism) patterns by ribosomal RNA and M13 probe. GC ratio and KNA homology values of th three isolates were approximately equal to those of ATCC type strain. The RFLP patterns by the rRNA and M13 probe showed some differences among C. thermocellum ATCC 27405, wild type strains and Clostridium thermohydrosulfuricum ATCC 33223, indicating that the two probes can be useful in subspecies- and apecies-identification.

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Agrobacterium-mediated Transformation of the Winter Mushroom, Flammulina velutipes

  • Cho, Jung-Hee;Lee, Seung-Eun;Chang, Who-Bong;Cha, Jae-Soon
    • Mycobiology
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    • v.34 no.2
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    • pp.104-107
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    • 2006
  • Flammulina velutipes was transformed efficiently by Agrobacterium-mediated transformation system. The transformation frequency was about 16% with the gill tissues of the fungal fruiting body. Southern hybridization and genetic analysis suggest that the introduced DNA was inserted onto different locations of the fungal genome, and inherited stably to the next generation via basidiospores. Transformation or gene tagging with Agrobacterium T-DNA based vector should be useful for wide ranges of genetic or molecular biological studies of the mushroom.

Nucleotide Sequence of a Proteinase Inhibitor I Gene in Potato (감자에 존재하는 단백질분해효소 억제제 I 유전자의 염기서열)

  • 이종섭
    • Journal of Plant Biology
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    • v.32 no.2
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    • pp.67-78
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    • 1989
  • Hybridization of DNA isolated from leaves of Russet Burbank potato with tomato cDNA as a probe revealed the presence of about ten inhibitor 1 genes in the genome. Screening of a genomic library of Russet Burbank potato resulted in isolation of seven different genomic clones carrying inhibitor I genes. One of the genomic clones, clone 2, contained two EcoRI fragments of 3.4 and 1.8 kb in size, respectively, which were hybridized with the probe. The nucleotide sequence of parts of the hybridizing EcoRI fragments revealed that they contain a complete gene which codes for an open reading frame of 107 amino acids. It is interrupted by two intervening sequences of 502 and 493 bp, situated at the positions of codons 17 and 43, respectively, of the open reading frame. Putative regulatory sequences, TATAAA and CCACT, were found at the 5' flanking region. In addition, a copy of a 100 bp repeat found at a tomato inhibitor I gene was identified.

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Isolation of Proteinase Inhibitor II Genes from Potato (감자로부터 단백질분해효소 억제제 II 유전자의 분리)

  • 이종섭
    • Journal of Plant Biology
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    • v.32 no.2
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    • pp.79-87
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    • 1989
  • Southern hybridization of genomic DNAs with radioactively labeled cDNA of tomato proteinase inhibitor II revealed that proteinase inhibitor II proteins in potato plants are encoded by a family of about 10 related sequences. Screening of potato EcoRI genomic library with the cDNA resulted in isolation of 13 recombinant phage clones which carry 3 different genomic regions. Of these clones, clones 8, 18, and 39 were subjected to restriction mapping and subcloning. Further characterization of the subclones of clones 8, 18 and 39 indicated that two inhibitor II genes are present on a 8.0 kb EcoRI fragment of clone 8, one on 3.3 and 0.8 kb EcoRI fragments of clone 18 and two genes on a 13.5 kb EcoRI fragment of clone 39.

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Isolation of $\beta$-Lactamase Inhibitory Protein from Streptomyces exfoliatus SMF19 and Cloning of the Corresponding Gene

  • PARK, HYEON-UNG;KYE JOON LEE
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.369-374
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    • 1996
  • The ${\beta}$-lactamase inhibitory protein (BLIP) produced by Streptomyces exfoliatus SMF19 was purified(33 kDa) and the N-terminal amino acid sequence was determined as NH2-ATSVVAWGGNND. Genomic DNA library of S. exfoliatus SMF19 was constructed in pWE15 and recombinants harbouring the corresponding gene were selected by colony hybridization to the mixture of 36-mer oligonucleotide designed from the N-terminal amino acid sequence. The corresponding gene (bliX) was isolated on a 4-kb ApaI fragment of S. exfoliatus SMF19 chromosomal DNA and then sequenced. The bliX consisting of 1, 119bp encoded a mature protein with a deduced amino acid sequence of 342 residues and also encoded a 40-amino-acid signal sequence. No significant sequence similarity to bliX was found by pairwise comparison using various protein and nucleotide sequences.

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MEMS for Heterogeneous Integration of Devices and Functionality

  • Fujita, Hiroyuki
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.7 no.3
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    • pp.133-139
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    • 2007
  • Future MEMS systems will be composed of larger varieties of devices with very different functionality such as electronics, mechanics, optics and bio-chemistry. Integration technology of heterogeneous devices must be developed. This article first deals with the current development trend of new fabrication technologies; those include self-assembling of parts over a large area, wafer-scale encapsulation by wafer-bonding, nano imprinting, and roll-to-roll printing. In the latter half of the article, the concept towards the heterogeneous integration of devices and functionality into micro/nano systems is described. The key idea is to combine the conventional top-down technologies and the novel bottom-up technologies for building nano systems. A simple example is the carbon nano tube interconnection that is grown in the via-hole of a VLSI chip. In the laboratory level, the position-specific self-assembly of nano parts on a DNA template was demonstrated through hybridization of probe DNA segments attached to the parts. Also, bio molecular motors were incorporated in a micro fluidic system and utilized as a nano actuator for transporting objects in the channel.

Draft Genome of an AmpC-β-Lactamase Producing Serratia marcescens Isolate from Fresh farm Tomatoes in South Africa

  • Maike Claussen;Stefan Schmidt
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.309-313
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    • 2023
  • Here we report essential features of the draft genome of an AmpC-β-lactamase-producing bacterial isolate obtained from farm tomatoes in South Africa. The isolate designated strain Tom1 featured a genome of 4950426 bp with a G+C% of 59.83. It was identified as Serratia marcescens by ribosomal multilocus sequence typing (rMLST), digital DNA-DNA hybridization (dDDH), average nucleotide identity (ANI), and phylogenetic analysis using reference genomes. Its genome encoded an AmpC-β-lactamase (blaSST-1), an efflux pump providing tetracycline resistance (tet(41)), and an aminoglycoside acetyltransferase (aac(6')-Ic). Additionally, genes encoding proteins involved in prodigiosin biosynthesis and associated with adherence, biofilm formation, virulence, and pathogenicity were detected.

Effect of Housing Systems of Cage and Floor on the Production Performance and Stress Response in Layer (계사 사육 형태가 산란계의 생산성과 스트레스 반응에 미치는 영향)

  • Sohn, Sea-Hwan;Jang, In-Surk;Son, Bo-Ram
    • Korean Journal of Poultry Science
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    • v.38 no.4
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    • pp.305-313
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    • 2011
  • This study was conducted to investigate the effects of housing systems on the productivity and physiological response as stress indicators in White Leghorn chickens. The chickens subjected to the conventional cages had a significantly lower viability, hen-housed egg production, egg weight and body weight compared with those to the floor pens. However, the hens housed in the conventional cages had a shorter day of the first egg and a greater egg quality compared with those housed in the floor pens. In addition, this study was also investigated to identify biological markers for assessing the physiological response of chickens under stress conditions. As biological markers, the amount of telomeric DNA was analyzed by quantitative fluorescent in situ hybridization on the nuclei of cells. The DNA damage rate of lymphocytes was also quantified by the comet assay. The amount of telomeric DNA of the lymphocytes, kidney and spleen was significantly higher in the chickens under floor pens than those under conventional cages. The DNA damage also increased in chickens raised under conventional cages, as compared to the chickens under floor pens. As results, we conclude that the chickens housed in conventional cages have a greater stressful status than those housed in floor pens.

Cloning of hadA-like Sigma Factor Gene from Streptomyces coelicolor A3(2) (Streptomyces coelicolor A3(2)에서 hrdA유사 Sigma 인자 유전자의 클로닝)

  • Hahn, Ji-Sook;Cho, Eun-Jung;Roe, Jung-Hye
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.264-270
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    • 1994
  • A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

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Karyotype Analysis and Physical Mapping of rDNAs Using Bicolor-FISH in Tiarella polyphylla D. Don (헐떡이풀의 핵형분석과 Bicolor FISH를 이용한 물리적 지도 작성)

  • Kim, Soo-Young;Lee, Joong-Ku
    • Korean Journal of Plant Resources
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    • v.20 no.5
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    • pp.446-450
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    • 2007
  • Tiarella polyphylla D. Don(Saxifragaceae) is a perennial herb and distributed in China, Japan, Taiwan and Korea. Especially, it only grows in Ulleung island of Korea. It has been using for asthma, bruise and audition troubles with main components of some Triterpenoids and seven oleanolic Saponins. There is only known its chromosomal number rarely and cytogenetic study was not done. From this study, karyotype analysis and chromosomal localization of 5S and 45S rDNAs using bicolor-FISH(fluorescence in situ hybridization) were carried out. The somatic metaphase chromosome number was 2n=2x=14 and the size of chromosomes ranged $1.66{\sim}3.50{\mu}m$. The chromosome complement consisted of four pairs of submetacentrics(chromosomes 1, 2, 3 and 6), two pairs of subtelocentrics(chromosomes 5 and 7) and one pair of telocentrics(chromosome 4). We also observed NOR(nucleolus organizer region) on the chromosome 4. In bicolor-FISH, one pair of 55 and 45S rDNA sites was detected on the centromeric region of chromosome 3 and short arm of chromosome 4, respectively. Bicolor FISH was very useful tool for the localization and identification of rDNAs on the chromosomes in Tiarella polyphylla.