• Title/Summary/Keyword: endonuclease

검색결과 247건 처리시간 0.025초

Comparison in Restriction Profile Analysis of Vibrio furnissi, Vibrio fluvialis, and Vibrio parahaemolyticus Bacteriophage from Sea Product

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • 제1권2호
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    • pp.99-103
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    • 1997
  • The bacteriophages lytic for Vibrio furnissi, Vibrio fluvialis and Vibrio parahaemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophages was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. furnissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, I with Bam HI and 2 with EcoR 1. V fluvialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR 1. V. parahamolyticus produced 13 sites with Hind III and 4 sites with EcoR 1. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V. furnissi phage were digested with 5 different restriction enzymes.

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TALENs Construction: Slowly but Surely

  • Hegazy, Wael Abdel Halim;Youns, Mahmoud
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3329-3334
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    • 2016
  • Cancer is thought to be a direct result of transcriptional misregulation. Broad analysis of transcriptional regulatory elements in healthy and cancer cells is needed to understand cancer development. Nucleases regulatory domains are recruited to bind and manipulate a specific genomic locus with high efficacy and specificity. TALENs (transcription activator-like effector nuclease) fused to endonuclease FokI have been used widely to target specific sequences to edit several genes in healthy and cancer cells. This approach is promising to target specific cancer genes and for this purpose it is needed to pack such TALENs into viral vectors. There are some considerations which control the success of this approach, targeting appropriate sequences with efficient construction of TALENs being crucial factors. We face some obstacles in construction of TALENs; in this study we made a modification to the method of Cermk et al 2011 and added one step to make it easier and increase the availability of constructs.

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

  • 김희선;엄수정;조쌍구;최용진
    • 한국미생물·생명공학회지
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    • 제22권6호
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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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PCR 기법을 이용한 Mycoplasma gallisepticum의 검출 (Detection of Mycoplasma gallisepticum using Polymerase Chain Reaction(PCR))

  • 이영주;김기석;김종완;탁연빈
    • 대한수의학회지
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    • 제39권1호
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    • pp.90-95
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    • 1999
  • A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

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

  • 강정훈;조영동
    • Journal of Ginseng Research
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    • 제9권2호
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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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Crystal structure of mismatch repair protein MutS and its complex with a substrate DNA

  • Ban, Changill
    • 한국결정학회:학술대회논문집
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    • 한국결정학회 2003년도 춘계학술연구발표회
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    • pp.16-16
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    • 2003
  • Mismatches in a DNA duplex are mainly due to DNA duplication errors that are generated by improper function of DNA polymerase. MutS, MutL and MutH are crucial proteins for the initiation of the methyl-directed mismatch repairing in bacteria. MutS has an ATPase activity md recognize the mismatched or unpaired bases on DNA. After binding to a mismatch, MutS recruits MutL to mediate the activation of MutH an endonuclease, which cleaves the 5' site of d(GATC) on the un-methylated strand. Both MutL and MutS also have essential roles in the subsequent removal and re-synthesis of the daughter strand. We have determined the crystal structures of either intact or active fragments of each of these proteins, both alone and complexed with ligands (DNA, ADP and ATP). The biochemical and mutagenesis studies based on the detailed 3-D structures led to new insights into the role of the ATPase activity of MutS in the mismatch recognition and directions for future investigation of mismatch repair.

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Identification of a Deoxyribonuclease I Inhibitor from a Phage-Peptide Library

  • Choi, Suk-Jung;Sperinde, Jeffrey J.;Szoka, Francis C. Jr.
    • Molecules and Cells
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    • 제19권1호
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    • pp.54-59
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    • 2005
  • Deoxyribonuclease I (DNase I) is a divalent cation dependent endonuclease and thought to be a significant barrier to effective gene delivery. The only known DNase I-specific inhibitor is monomeric actin which acts by forming a 1:1 complex with DNase I. Its use, however, is restricted because of tendency to polymerize under certain conditions. We screened two random phage peptide libraries of complexity $10^8$ and $10^9$ for DNase I binders as candidates for DNase I inhibitors. A number of DNase I-binding peptide sequences were identified. When these peptides were expressed as fusion proteins with Escherichia coli maltose binding protein, they inhibited the actin-DNase I interaction ($IC_{50}=0.1-0.7{\mu}M$) and DNA degradation by DNase I ($IC_{50}=0.8-8{\mu}M$). Plasmid protection activity in the presence of DNase I was also observed with the fusion proteins. These peptides have the potential to be a useful adjuvant for gene therapy using naked DNA.

Lactobacillus casei의 phospho-$\beta$-galactosidase 유전자의 대장균내 분자클로닝 (Molecular cloning of phospho-$\beta$-galactosidase gene of lactobacillus casei in escherichia coli)

  • 문경희;박정희;최순영;이유미;김태한;김연수;민경희
    • 미생물학회지
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    • 제27권3호
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    • pp.188-193
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    • 1989
  • Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

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RPA-governed Endonuclease Switching during Eukaryotic Okazaki Fragment Processing.

  • Bae, Sung-Ho;Bae, Kwang-Hee;Kim, Jung-Ae;Seo, Yeon-Soo
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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    • pp.22-22
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    • 2001
  • At the eukaryotic replication fork, discontinuous synthesis of lagging strand DNA gives rise to Okazaki fragments carrying ribonucleotides derived from the primer RNA at their 5' ends. Efficient removal of these ribonucleotides is vital for maintaining genome integrity. In this report we show that the endonucleases Dna2 and Fen1 act sequentially to facilitate the complete removal of the primer RNA.(omitted)

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

  • 이정헌;임용표;최광태
    • Journal of Ginseng Research
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    • 제17권1호
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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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