• Title/Summary/Keyword: Tn5 polypeptides

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Amplified synthesis and stability of Tn5 polypeptides in escherichia coli (대장균에서의 Tn5 단백질 증폭생합성 및 안정성)

  • 정재성;정재훈
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.323-333
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    • 1989
  • Plasmid DNA molecules containing strong promoter upstream from IS50L or IS50R, the two insertion sequences that flank Tn5, were constructed to amplify the synthesis of Tn5-encoded polypeptides. When proteins made by cells that contain these plasmids were analyzed on polyacrylamide gels, enhanced synthesis of IS50R polypeptides could be detected. Synthesis of this polypeptide apparently is initiated within the large open reading frame of this element. In addition, the stability of IS50L-and IS50R-encoded polypeptides was analyzed. It was found that IS50L polypeptides are relatively unstable in vivo. This instability could account for the observed inability of this element to promote transposition.

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Genetic Organization of the Recombinant Bacillus pasteurii Urease Genes Expressed in Escherichia coli

  • Kim, Sang-Dal;Hausinger, Robert P.
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.108-112
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    • 1994
  • The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major ($M_r$ 67,000) and minor ($M_r$ 20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.

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Fractionation of Angiotensin Converting Enzyme(ACE) Inhibitory Peptides from Soybean Paste (된장으로부터 Angiotensin Converting Enzyme(ACE) 저해 Peptide의 분획)

  • Shin, Zae-Ik;Ahn, Chang-Won;Nam, Hee-Sop;Lee, Hyung-Jae;Lee, Hyung-Joo;Moon, Tae-Hwa
    • Korean Journal of Food Science and Technology
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    • v.27 no.2
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    • pp.230-234
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    • 1995
  • Angiotensin converting enzyme(ACE) inhibitory peptides lowering blood pressure were fractionated from a commercial soybean paste(Doenjang). When the freeze-dried sample of soybean paste was extracted with cold water, the recovery yield of total nitrogen(TN) was shown to be 73.3% in 30 minutes. The cold water extract was filtered through PM-10 membrane(Amicon) for 3 hours in order to remove high molecular weight polypeptides. The TN and salt of ultrafiltrate were recovered to 80.8% and 99.2%, respectively, and its ACE $IC_{50}$ was $41.8{\mu}g/ml$. When the ultrafiltrate was divided into 7 fractions by reverse phase prep-HPLC, F5 fraction showed the highest ACE inhibitory activity ($IC_{50}=6.8{\mu}g/ml$) and salt could be collected into F1 fraction. Subsequently, the F5 fraction was divided into another five fractions by ion exchange prep-HPLC, all of which appeared to be high ACE inhibitory activity($IC_{50}=2.5{\sim}8.3{\mu}g/ml$). Among them, F53 fraction had the highest ACE inhibitory activity, and its main amino acid component was found to be histidine.

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