• Title/Summary/Keyword: Restriction endonuclease

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Complementary DNA Cloning and Restriction Mapping of Nuclear Inclusion Body and Coat Protein Genes of Turnip Mosaic Virus-Ca Strain Genomic RNA (순무모자이크 바이러스 Ca계통 핵봉입체와 외피단백질 유전자의 cDNA 클로닝 및 제한효소 지도작성)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.235-239
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    • 1994
  • Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.93.2-97
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    • 1992
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnificus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio pnrahemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios. Key words . plasmid, Vibrio anguillarum, Vibrio fluvialis, Vibrio uulnincus, Vibrio mimicus, Vibrio furnissi, Vibrio parahemolyticus.

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A Micromethod for Rapid and Simple Isolation of Genomic DNA from Small Scale Culture of Bifidobacterium (소량의 Bifidobacterium 배양액에서 genomic DNA 추출을 위한 신속/간단한 방법)

  • Jeakal, Soo;Park, Hee-Kyung;Song, Ji-Eun;Heo, Tae-Ryeon;So, Jae-Seong
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.781-783
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    • 1995
  • A method is described for the rapid and simple isolation of genomic DNA from 3 ml culture of Bifidobacterium. The method is expected to be used in gene manipulation of Bifidobacterium spp. The isolated DNA using this method is shown to be an excellent substrate for restriction endonuclease digestion and ligation with T4 DNA ligase.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.93-97
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    • 1997
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnficus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio parahaemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios.

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Cloning of Steroid $\Delta^1$-dehydrogenase Gene of Arthrobacter simplex IAM 1660

  • Bae, Moo;Bae, Song-Mee;Lee, Mi-Kyung;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.142-144
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    • 1996
  • To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.

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Characterization of Endogeneous Plasmids from Two Bacillus Isolates (Bacillus 속 분리균 2종의 내재형 Plasmids 특성분석)

  • 윤기홍
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.364-369
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    • 1999
  • In order to obtain the suitable plasmids for constructing plasmid vectors of Bacillus species, endogeneous plasmid DNAs were screended from thermo-tolerant soil bacteria. Based on agarose gel electrophoresis patterns of the isolated plasmid DNAs, two strains harboring small-size plasmids were selected. The isolated were identified to belong to the genus Bacillus on the basis of their morphological and biochemical properties, and named Bacillus sp. 3-3 and 77-8, respectively. The restriction endonuclease maps were determined for four plasmids including two plasmids from each Bacillus isolates. It is interesting that Bacillus sp. 3-3 and 77-8 have an identical plasmid according to the restriction maps. The three kinds of hybrid plasmids constructed by introducing each plasmid of two isolates into a Escherichia coli plasmid vector. pUCCm18 containing chloramplenicol resistance gene active in Bacillus strains, could be replicated in B. subtilis and B. licheniformis. These plasmids are very stable in B. subtilis, suggesting that the Bacillus plasmids identified in this work would be useful for development of new cloning vectors for Bacillus strains.

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Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene(estII) of Bacillus Stearothermophilus in Escherichia coli (Bacillus stearothermophilus Acetyl Exterase 유전자(estII)의 클로닝과 Escherichia coli에서의 발현)

  • Kim, Hee-Sun;Eom, Soo-Jung;Cho, Ssang-Goo;Choi, Yong-Jin
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.599-606
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    • 1994
  • Bacillus stearothermomophilus, a strong xylan degrader, was confirmed to express multiple esterase activities in addition to the major xylanolytic enzymes. One of the genes encoding the esterases was isolated from the genomic library of B. stearothermophilus constructed with EcoRl restriction endonuclease and pBR322 plasmid. Three recombinant plasmids showing the tributyrin degrading activity were selected from approximately 7, 000 E. coli HB101 transformants, and were found to have the same insert of a 3.2 kb DNA fragment. Restriction mapping and hybridization studies revealed that the gene(estII) on the hybrid plasmid (pKMG7) had originated from the B. stearothermophilus chromosome, and was distinct from the estl, another esterase gene of B. stearothermophilus isolated in the previous work. The E. coli cells harboring pKMG7 produced an acetylxylan esterase that exibited similar substrate specificity to the esterase encoded by the estI gene.

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Effect of Ginseng Saponin Fraction on Cleavage of pBR322 by Several Restriction Endonuclease (인삼 Saponin분획이 제한효소에 의한 pBR322 절단에 미치는 영향)

  • 강정훈;조영동
    • Journal of Ginseng Research
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    • v.9 no.2
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    • pp.240-247
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    • 1985
  • Attempts were made to see if we could cut more pBR322 in the presence of ginseng saponin fraction in connection with possibly for shortening the enzymatic reaction time and the amount of the enzyme to be used. The following results were obtained restriction endonucleases such as AccI, XhoII, SaII, and HincII were observed to cut pBR322 efficiently at 10-1% ginseng saponin fraction. In case of BamHI, 10-2% ginseng saponin fraction was observed to the most effective concentration. Such cumulative results suggest that ginseng saponin fraction would play important role as far as the cleavage of pBR322 for short period by endonucleases is concerned.

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Isolation and Characterization of Chloroplast DNA in Korea Ginseng, Panax gindeng C.A. Meyer (고려인삼(Panax ginseng C.A. Meyer)의 엽록체 DNA 분리 및 특성조사)

  • Lee, Jeong-Heon;Lim, Yong-Pyo;Choi, Kwang-Tae
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.39-44
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    • 1993
  • In Korean ginseng, Panax ginseng C.A Meyer, it was difficult to isolate chloroplast DNA with classical methods, because of the high polysaccharide content of ginseng chloroplast The simple and efficient method of chloroplast DNA isolation from ginseng leaves has been developed by motificalion of recently advanced methods. Also, it can be successfully applied to ctDNA isolation of Chinese cabbage, radish, petunia tobacco as well as ginseng. Isolated chloroplast DNA from ginseng was digested with various restriction endonucleases. It was estimated that the molecular weight of Korean ginseng chloroplast DNA was about 142 kb. There was no difference in restriction endonuclease digestion patterns between two variants of Korean ginseng, which are Jakyung-Jong (violet-stem variant) and Hwang- sook-Jong (yellow-berry variant).

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Cloning and Expression of the Bdi Methylase Gene in E. coli (대장균 내에서의 Bdi I Methylase 유전자의 클로닝과 발현)

  • 전희숙;김용석;최경래;노현모
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.40-45
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    • 1987
  • The gene for the Bdi I modification enzyme, which is one of Bdi I restriction-modification system, fromBrevibacterium divaricatum FERM 5948 was cloned and expressed in E. coli. For cloning of the Bdi I methylase gene, we have initially used three cloning site(EcoRI, BamHI and Sal I) of plasmid vector pBR 322 and adopted the retransformation method after Bdi I restriction endonuclease cleavage. Selection of transformants carrying the gene was based on the resistance of the modified plasmid encoding the enzyme to cleavage by Bdi I restriction enzyme, and the recombinant plasmid pBDIM 116 containing 5.6kb EcoRI insery was proved to carry the gene. Crude cell extracts prepared from strains carrying the plasmid pBDIM 116 contained an S-adenosylmethionine-dependent methyltransferase activity specific for the Bdi I recognition site, ATCGAT. The restriction map was constructed with 11 restriction enzyme, and the Bdi I restriction-modification system was also discussed.

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