• 제목/요약/키워드: Hind III

검색결과 207건 처리시간 0.024초

생굴로부터 bacteriophage의 분리 및 부분특성 (Isolation and Partial Characterization of Bacteriophage from Oyster)

  • 김영희
    • 한국환경과학회지
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    • 제5권5호
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    • pp.605-610
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    • 1996
  • 시판중인 생굴을 대상으로 Vibrio 속을 숙주로 하는 bacteriophage의 분리를 시도하였다. 4 종의 Vibrio 속과 5 혈청형의 Vibrio parahaemolytius를 실험대상으로 한 결과 배brio parahaemolyticus 2 혈청형에서만 bacteriophage가 분리되었다. 분리된 phage의 plaque 크기는 0.4mm였으며 전자현미경적 형태는 미부가 뚜렷하지 않은 육각형의 두부가 관찰되었고 크기는 67nm$\times$83nm 였으며, PFV/ml은 1.25$\times$$10^{11}$이었다. 분리된 phage는 chloro-form에 감수성을 나타내었다. 분리된 phage의 genomic 특성을 규명하기 위하여 핵산을 분리한 결과 DNA로 판명되어졌으며 두 혈청형 모두 제한효소 처리한 결과 Eco R I으로 4부위의 절단양상과 Hind III로부터 14부위 절단양상이 관찰되었다.

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Linkage of the Kanamycin Resistance Gene with the Streptothricin Resistance Gene in Staphylococcus aureus SA2

  • Shin, Chul Kyo;Sung Hwan Im;Woo Koo Kim;Kyung Bo Moon
    • Journal of Microbiology and Biotechnology
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    • 제6권3호
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    • pp.219-220
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    • 1996
  • The pKH2 isolated from the multidrug-resistant Staphylococcus aureus SA2 is a 40.98-kb plasmid and mediates resistance to ampicillin, clindamycin, erythromycin, kanamycin, and streptomycin. The 3.4-kb HindIII fragment conferring kanamycin resistance was cloned from the pKH2 into pBluescriptII $KS^+$ and partial sequence determination of that fragment was carried out. Sequence analysis revealed that the kanamycin resistance gene which encoded aminoglycoside 3'-phosphotransferase was linked to the streptothricin resistance gene. But a nonsense mutation was found in the streptothricin resistance gene and this mutation resulted in a truncated protein of streptothricin acetyltransferase. Homology comparison with nucleotide sequence databases revealed that the 3.4-kb HindIII fragment of pKH2 had been derived not from S. aureus but from Gram-negative Campylobacter coli.

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Cloning and Characterization of a Gene Encoding 22 kDa Functional Protein of Bacteriophage MB78

  • Gupta, Lalita;Chakravorty, Maharani
    • BMB Reports
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    • 제38권2호
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    • pp.161-166
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    • 2005
  • Functional protein of MB78 bacteriophage having apparent molecular weight of 22 kDa is expressed from 1.7 kb HindIII G fragment. The nucleotide sequence of this fragment showed two open reading frames of 222 and 196 codons in tail-to-tail orientation separated by a 62-nucleotide intercistronic region. The ORF of 22 kDa protein is present in opposite orientation, i.e. in the complementary strand, preceded by a strong ribosomal binding site and a promoter sequence. Another ORF started from the beginning of the fragment whose promoter region and translational start site lies in the 0.45 kb HincII U fragment which is located next to the HindIII G fragment, that has the sequence for DNA bending. 3' end of the fragment has high sequence homology to the EaA and EaI proteins of bacteriophage P22, a close relative of MB78 phage.

Molecular Cloning of $\beta$-Galactosidase from Bacillus subtilis HP-4

  • Kim, Jeong-Ho;Lee, Jae-Chang;Huh, Jeong-Won;Chung, Ki-Chul
    • Journal of Microbiology and Biotechnology
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    • 제1권4호
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    • pp.227-231
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    • 1991
  • A gene coding for a $\beta$-galactosidase of Bacillus subtilis HP-4 was cloned in E. coli JM109 by inserting HindIII digested fragment of B. subtilis HP-4 chromosomal DNA into the site of pBR322 and selecting recombinant transformant showing blue color on X-gal plate. The recombinant plasmid, named pBG109, was found to contain the 1.4 Kbp HindIII fragment originated from B. subtilis HP-4 chromosomal DNA by Southern hybridization. The cloned gene was stably maintained and expressed in E. coli JM109 and the pBG109 encoded $\beta$-galactosidase had the same enzymatic properties as those of $\beta$-galactosidase produced by B. subtilis HP-4.

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Molecular Cloning and Expression of a Xylanase Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Kang, Yun-Sook;Park, Young-Seo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제1권4호
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    • pp.251-255
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    • 1991
  • A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

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Bacillus stearothermophilus Acetyl Xylan Esterase 유전자의 크로닝과 Escherichia coli에서의 발현 (Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene of Bacillus stearothermophilus in Escherichia coli)

  • 김인숙;조쌍구;최용진
    • 한국미생물·생명공학회지
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    • 제21권6호
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    • pp.542-548
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    • 1993
  • Bacillus stearothermophilus was shown to express multiple xylanolytic enzymes including acetyl xylan esterase. Genomic DNA of the strain partially digested with HindIII was ligated into the HindIII site of pBR322, and expressed in E. coli HB101 cells in order to clone the gene for acetyl xylan esterase. One transformant among 4000 screened formed a clear zone around its colony on the LB agar supplemented with 1.0% tributyrin. The functional clone harbored the recombinant plasmid pKMG5 with an insert of 5.1kb.

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Genetic Polymorphisms of t-PA and PAI-1 Genes in the Korean Population

  • Kang, Byung-Yong;Lee, Kang-Oh
    • Animal cells and systems
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    • 제7권3호
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    • pp.249-253
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    • 2003
  • Abnormalities in fibrinolysis system is associated with risk of hypertension. In this report, the Alu repeat insertion/deletion (I/D) polymorphism of tissue plasminogen activator (t-PA) and the Hind III RFLP of plasminogen activator inhibitor-1 (PAI-1) genes were investigated in 115 normotensives and 83 patients with hypertension, and their association with anthropometrical data and plasma biochemical parameters were analyzed. There were no significant differences in the gene frequencies of the two candidate genes between normotensives and hypertensives, respectively. Our results indicate lack of associations between the two polymorph isms in t-PA and PAI-1 genes and risk of hypertension in the population under study. However, the Hind III RFLP of PAI-1 gene was significantly associated with plasma glucose level, suggesting its role in glucose metabolism. It needs to be tested whether this RFLP of PAI-1 gene is associated with insulin resistance syndrome or non-insulin dependent diabetes mellitus (NIDDM) in the Korean population.

미토콘드리아 DNA 제한효소 절단부위 변이에 의한 Anopheles quadrimaculatus (Say) 모기의 자매종 구별 (restriction Site Polymorphism of mtDNA for differentiating Anopheles quadrimaculatus (Say) Sibling Species)

  • 김상석
    • 한국응용곤충학회지
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    • 제29권2호
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    • pp.132-135
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    • 1990
  • Anopheles quadrimaculatus( Say) 자매종 간의 미토콘드리아 DNA의 제한효소 절단부위변이를 Aedes albopictus의 마토콘드리아 cDNA를 probe로 이용하여 조사하였다. DNA hybridization에 의해 두 속간에는 mtDNA 영기서열의 상당한 상동성이 있음을 알 수 있었다. 개체 모기로부터 분리한 DNA를 제한효소를 사용하여 절단한 결과 자매종간에 다른 양상을 볼 수 있었으며 Hind III에 의한 mtDNA 절편만으로도 자매종들을 동정할 수 있었다.

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Fuarium oxysporum의 유연관계 분석을 위한 Recombinant DNA의 Probe로서의 이용 가능성 (Potential Applications of Recombinant DNA Probes for Relatedness Analysis of Fusarium oxysporum)

  • 김홍기;김영태;유승헌
    • 한국식물병리학회지
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    • 제10권1호
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    • pp.1-6
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    • 1994
  • Randomly chosen recombinant clones of Fusarium oxysporum were analysed to select useful probes for relatedness analysis of Fusarium oxysporum. Genomic DNA of F. oxysproum f. sp. cubense, digested with HindIII, was ligated to pUC118 and used to transform Escherichia coli strai DH5$\alpha$. Three clones were identified that hybridized to mutiple restriction fragments of some formae speciales of F. oxysporum. These probes detected repetitive sequences in HindIII or EcoRI digested DNAs. Repeated copy clone pFC46, pFC52 and pFC54 showed evident polymorphisms among ten formae speciales of this fungus. Since clone pFC 52 strongly hybridized to multiple EcoRI-digested restriction fragments of f. sp. cubense, it may be useful as a probe for analysis of other genetic characteristics of this forma specialis. The results suggest that our clones might be very useful as probes for relatedness analysis between or within formae speciales of Fusarium oxysporum.

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Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현 (Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli)

  • 정영철;김양우;노종수;성낙계;강신권
    • 한국미생물·생명공학회지
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    • 제18권6호
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    • pp.633-639
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    • 1990
  • Cellulase 복합체와 xylanase를 동시에 분비하는 Pseudomonas sp. LBC 505와 CYC 10의 cellulase 유전자를 pUC19를 사용하여 E.coli에 클로닝시켰다. Congo red 염색시 노란색 환을 형성하는 대장균 형질전환에서 7.0Kb-와 4.6Kb-HindIII 단편을 함유한 재조합 플라스미드 pLC1과 pLC2를 가각 분리하였다. DNA hybridization 실험에서 pLC1 과 pLC2는 Pseudomonas sp. LBC 505와 CYC 10 유래임이 각각 밝혀졌고, Immunoassay 실험에서도 유사성이 인정되었다. pLC1을 함유하고 있는 대장균은 cellulas의 24를 세포외로 분비하였고, 효소활성은 모균주에 비해 1.4배 증가하였다. pLC1과 pLC2의 효소학적 성질도 모균주와 동일하였으며, 기질특이성과 HPLC로 유리당을 분석한 결과, 클로닝된 유전자는 endo type인 것으로 나타났다.

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