• Title/Summary/Keyword: Nuclease S1

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Study on the Specificity Alteration of Mammalian UV Endonuclease III

  • Lee, Jae-Yung;Kim, Joon
    • BMB Reports
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    • v.30 no.1
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    • pp.66-72
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    • 1997
  • A mammalian DNA repair enzyme, UV endonuclease III which also functions as a ribosomal protein S3 (rpS3), was purified from mouse cells and characterized. UV endonuclease III was previously cloned and known to yield a peptide of 32 kDa upon expression in E. coli [Kim et al., (1995) J. Bioi. Chem. 270, 13620-13629]. However, biochemically purified UV endonuclease III, which has a sedimentation coefficient of 3.25, appears to have an additional peptide of 28 kDa. It appears that two bands were derived from one complex, judging from the comparison of the nuclease activity on the native and SDS-gel electrophoreses. UV endonuclease III becomes non-specific upon purification and this phenomenon is more significant in the case of pure fractions of the enzyme. Non-specific activity was not influenced by pH or any salt conditions.

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Restriction pattern of the nucleic acid of Synechococcus sp. cyanophage

  • Park, Jong-Geun;Kim, Min;Choi, Yong-Keel;Yoon, Sung-Nyo
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.1-6
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    • 1996
  • The nucleic acid of Synechococcus sp. cyanophage was identified as double-stranded DNA by the result of digestion with enzymes such as exonucleases, DNase, and S1 nuclease, and by acridine orange staining. The cyanophage DNA was cleaved with several restriciton ehdonucleases such as ApaI, BamHI, Bg/II, HaeIII, Eco RI, HindIII, PstI, AND aPAI gave the clearest sets of bands on agarose gels and the fragment numbers for each were 12, 20, 29, 20, and 7, respectively. The sums of the size from Bam HI and PstI digestions were estimated approximately 227$\pm$4 kb, which are in agreement with the result of the pulsed field gel electrphoresis. This virus is thought to have the largest genosome among those of known cyanophages, which corresponds to the largest haed ot 90 nm when compared with the head sizes of cyanophages discovered since 1963.

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Secondary Structure for RNA Aptamers Binding to Guanine-Rich Sequence in the 5'-UTR RNA of N-Ras Oncogene

  • Cho, Bongrae
    • Journal of the Korean Chemical Society
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    • v.65 no.2
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    • pp.121-124
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    • 2021
  • RNA molecules which bind to the G-rich sequence in the 5'-UTR RNA which plays an important role in expression of N-ras, were selected. The secondary structures of five selected RNA aptamers including primer sequence were found by the CLC RNA workbench ver. 4.2 program (www.clcbio.com) and investigated with RNA structural probes such as RNase T1 which has specificity for a G in single-stranded region, RNase V1 specific for double strand and nuclease S1 specific for single strand. The generalized secondary structure model was proposed and characterized. It was composed of a central long double strand region flanked by single strand region at both end sides. The double strand region had an internal single-strand region and bulges. The single strand loop in the right side was composed of four or five nucleotides.

Identification of Differentially Expressed Genes by TCDD in Human Bronchial Cells: Toxicogenomic Markers for Dioxin Exposure

  • Park, Chung-Mu;Jin, Kyong-Suk;Lee, Yong-Woo
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.1-9
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    • 2012
  • Differentially expressed genes by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were identified in order to evaluate them as dioxin-sensitive markers and crucial signaling molecules to understand dioxin-induced toxic mechanisms in human bronchial cells. Gene expression profiling was analyzed by cDNA microarray and ten genes were selected for further study. They were cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1), S100 calcium binding protein A8 (calgranulin A), S100 calcium binding protein A9 (calgranulin B), aldehyde dehydrogenase 1 family, member A3 (ALDH6) and peroxiredoxin 5 (PRDX5) in up-regulated group. Among them, CYP1B1 was used as a hallmark for dioxin and sharply increased by TCDD exposure. Down-regulated genes were IK cytokine, interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), nuclease sensitive element binding protein 1 (NSEP1), protein tyrosine phosphatase type VI A, member 1 (PTP4A1), ras oncogene family 32 (RAB32). Although up-regulated 4 genes in microarray were coincided with northern hybridization, down-regulated 5 genes showed U-shaped expression pattern which is sharply decreased at lower doses and gradually increased at higher doses. These results introduce some of TCDD-responsive genes can be sensitive markers against TCDD exposure and used as signaling cues to understand toxicity initiated by TCDD inhalation in pulmonary tissues.

Studies on Staphylococci Isolated from Bovine Udder Infections I. Biochemical Properties of Staphylococcus aureus (젖소유방염유래(乳房炎由來) 포도구균(葡萄球菌)에 관한 연구(硏究) I. Staphylococcus aureus의 생화학적특성(生化學的特性))

  • Park, Cheong-Kyu
    • Korean Journal of Veterinary Research
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    • v.22 no.1
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    • pp.15-21
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    • 1982
  • The biochemical properties of strains of Staphylococcus aureus isolated from cattle with peracute gangrenous mastitis, acute systemic mastitis and chronic mastitis were examined. Of 261 strains of Staphylococci isolated from quarters with clinical mastitis, 140 (53.6%) were classified as Staphylococcus aureus and 121 (46.4%) were coagulase-negative staphylococci. All the strains of Staphylococcus aureus isolated from quarters with peracute gangrenous mastitis and acute systemic mastitis showed production of alpha heamolysin, coagulase, lipase, phosphatase, nuclease and gelatinase, and fermentation of mannitol. However, of 114 strains of Staphylococcus aureus obtained from quarters with chronic mastitis, 64 (56.1%) possessed alpha lysin, and 43 (37.7%) produced lipase in egg yolk medium. The most common hemolytic pattern of the strains associated with chronic mastitis was ${\beta}{\delta}$ type.

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Production of Yeast Extract by a Combined Method of Autolysis and Enzymatic Hydrolysis (자기소화와 효소가수분해 방법을 병용한 효모 추출물의 제조)

  • 인만진;채희정
    • KSBB Journal
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    • v.19 no.4
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    • pp.245-249
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    • 2004
  • A combined method of autolysis and enzymatic hydrolysis of baker's yeast was developed for the production of yeast extract, which is widely used as a natural food ingredient. From statistical analysis, NaCl and ethanol addition were found to be significantly effective factors in autolysis of yeast. The optimum dosages of salt and ethanol were 3% and 1%, respectively. Heat treatment and the use of cell lytic enzyme were not significantly effecting on the autolysis. Yeast hydrolysate was prepared by autolysis, followed by enzymatic hydrolysis using proteases, nuclease and deaminase. Additionally, the hydrolysate was processed by downstream process including Maillard reaction and debittering. The total dry matter yield and total nitrogen yield for the process were 76% and 59%, respectively. Compared to a process using brewer's yeast, when baker's yeast was used as a raw material, a higher recovery yield was obtained.

Role of Chromatin Structure in HMRE Mediated Transcriptional Repression of the HSP82 Heat Shock Gene

  • Lee, See-Woo;Gross, David S.
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.40-48
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    • 1996
  • We have examined the chromatin structure of the HMRE/HSP82 and HMRa/HSP82 allels using three complementary approaches : DNase I chromating footprinting, micrococcal nuclease (MNase) nucleosome-protected ladder assay, and an in vivo E. coli dam methylase accessibility assay. The footprinting results indicate that the promoter and silencer sequences are assembled into nucleoprotein complexes which exhibit no detectable change in structure, despite a 70-fold range in expression levels. In addition, the promoter region of the HMRa/HSP82 allele is cleaved randomly by MNase in all cases, indicating the absence of anonical nucleosomes over this region irrespective of SIR4 or heat-shock. Finally, no discernible difference in the accessibility of the HMRE/HSP82 locus to dam methylase in SIR4 vs. sir4 cells was seenm which again suggests that the chromatin structure of HMRE/HSP82 allele is identical regardless of SIR4. Altogether, our results indicate that in contrast to other observations of the silent mating-type loci, no discernible structural alteration is detected at either HMR/HSP82 allele regardless of SIR genetic background or transcriptional state of the gene.

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Nucleotide Sequence and Analysis of a Xylanase gene (xynS) from Alkali-tolerant Bacillus sp. YA-14 and Comparison with Other Xylanases

  • Yu, Ju-Hyun;Park, Young-Seo;Yum, Do-Young;Kim, Jin-Man;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.139-145
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    • 1993
  • The nucleotide sequence of the xylanase gene (xynS) from alkali-tolerant Bacillus sp. YA.14 was determined and analyzed. A 639 base pairs open reading frame for xynS gene was observed and encoded for a protein of 213 amino acids with a molecular weight of 23, 339. S1 nuclease mapping showed that the transcription initiation site of the xynS gene did not exist in the cloned DNA. Ribosome binding site sequence with the free energy of -18.8 Kcal/mol was observed 8 base pairs upstream from the initiation codon, ATG. The proposed signal sequence consisted of 28 amino acids, of which 3 were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YA-14 xylanase showed 48% homology with Bacillus sp. YC-335 xylanase and 96% homology with xylanases from B. subtilis and B. circulans.

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Sequence Analysis and Expression of Xylanase Gene (xynY) from Alkalophilic Bacillus sp. YC-335

  • Park, Young-Seo;Yum, Do-Young;Kim, Jin-Man;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.224-231
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    • 1993
  • The nucleotide sequence of the xylanase gene (xynY) from alkalophilic Bacillus sp. YC-335 was determined and analyzed. An open reading frame of 1, 062 base pairs for xynY gene was observed and encoded for a protein of 354 amino acids with a molecular weight of 38, 915. S1 nuclease mapping showed that the transcription initiation sites of the xynY gene were different in Bacillus sp. YC-335 and Escherichia coli HB101 (pYS55). S1 mapping also showed that -10 region of the xynY gene recognized by RNA polymerases of E. coli and Bacillus sp. YC-335 were TACAGT and TATGAT , respectively. A ribosome binding site sequence with the free energy of -17.0 Kcal/mol was observed 9 base pairs upstream from the unusual initiation codon, TTG. The proposed signal sequence consisted of 27 amino acids, 2 of which were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YC-335 xylanase showed more than 50% homology with xylanases from B. pumilus, B. subtilis, and B. circulans.

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GSTP1, ERCC1 and ERCC2 Polymorphisms, Expression and Clinical Outcome of Oxaliplatin-based Adjuvant Chemotherapy in Colorectal Cancer in Chinese Population

  • Li, Hui-Yan;Ge, Xin;Huang, Guang-Ming;Li, Kai-Yu;Zhao, Jing-Quan;Yu, Xi-Miao;Bi, Wen-Si;Wang, Yu-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3465-3469
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    • 2012
  • Aim: Platinum agents have shown to be effective in the treatment of colorectal cancer. We assessed whether single nucleotide polymorphisms (SNPs) in GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln might predict the overall survival in patients receiving oxaliplatin-based chemotherapy in a Chinese population. Methods: SNPs of GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln in 335 colorectal cancer patients were assessed using TaqMan nuclease assays. Results: At the time of final analysis on Nov. 2011, the median follow-up period was 37.7 months (range from 1 to 60 months). A total of 229 patients died during follow-up. Our study showed GSTP1 Val/Val (HR=0.44, 95% CI=0.18-0.98), ERCC1 C/C (HR=0.20, 95% CI=0.10-0.79) and ERCC2 G/G (HR=0.48, 95% CI=0.19-0.97) to be significantly associated with better survival of colorectal cancer. GSTP1 Val/Val, ERCC1 C/C and ERCC2 G/G were also related to longer survival among patients with colon cancer, with HRs (95% CIs) of 0.41 (0.16-0.91), 0.16 (0.09-0.74) and 0.34 (0.16-0.91), respectively. Conclusion: GSTP1, GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln genotyping might facilitate tailored oxaliplatin-based chemotherapy for colorectal cancer patients.