• Title/Summary/Keyword: Sp factor

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Characterization of a Cell Aggregation Factor from Aspergillus sp.LAM 94-142 (Aspergillus sp. LAM 94-142가 생산하는 세포응집물질의 특성)

  • 이동희;함동수
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.506-512
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    • 1995
  • A cell aggregation factor produced by Aspergillus sp. LAM 94-142 was purified and partially characterized. The factor was purified about 15 folds from culture broth by IRA 420 and IRC 120 treatment, 1% NaCl added acetone precipitation, and Sepharose 4B column chromatography with overall yield of 48%. It was heteropolysaccharide consisted of mannose, arabinose, and glucose with a molar ratio, 31:17:2, and its molecular weight was estimated to be about 900,000 daltons by Sepharodse 4B gel filtration method. The optimum pH and temperature was 8 and 40$\circ$C, respectively. The factor was stable in pH range of 3-9 and at 100$\circ$C for 90 min. The cell aggregation activity of the factor was inhibited by the addition of Hg$^{2+}$, Fe$^{2+}$, Cu$^{2+}$, and some polypeptides such as milk casein or hemoglobin. The factor aggregated Bacillus subtilis, B. macerans, B. turingiensis, E. coli, Peudomonas aeruginosa, P. fluorescens, P. malophilia, and weakly aggregated Staphylococcus sp., Sarcina lutea, P. putida and Cryptococcus neoformnans, but it didn't aggregate various strains of Candida sp. and Saccharomyces sp.

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Sp1 Decoy Oligodeoxynucleotides Inhibit Serum-induced Mesangial Cell Proliferation (Sp1 Decoy Oligodeoxynucleotides에 의한 사구체 혈관간세포 증식억제 효과)

  • Chae Young Mi;Kim Sung Young;Park Kwan Kyu;Chang Young Chae
    • KSBB Journal
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    • v.19 no.5
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    • pp.335-340
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    • 2004
  • Mesangial expansion caused by cell proliferation and glomerular extracellular matrix accumulation is one of the earliest renal abnormalties observed at the onset of hyperglycemia in diabetes mellitus. Transcription factor Sp1 is implicated in the transcriptional regulation of a wide range of genes participating in cell proliferation, and is assumed to play an essential role in mesangial expansion, transforming growth factor (TGF)-$\beta$1, plasminogen activator inhibitor (PAI)-1. We have generated a phosphorothioated double-stranded Sp1-decoy oligodeoxynucleotide that effectively blocks Sp1 binding to the promoter region for transcriptional regulation of TGF-$\beta$1 and PAI-1. The Sp1 decoy oligodeoxynucleotide suppressed transcription of these cytokines and proliferation of primary rat mesangial cells in response to serum stimulation. These results suggest that the Sp1 decoy oligodeoxynucleotide could bea powerful tool in preventing the pathogenesis of renal hypertrophy.

A Study on the Manganese Oxidation and Characteristics of Aeromonas sp (Aeromonas sp. MN44의 특성과 망간 산화에 관한 연구)

  • Koo Jong Seo;Park Kyeong Ryang
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.94-99
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    • 2005
  • Sixty four bacterial colonies which were able to oxidize the manganese were isolated from soil samples in Mokcheon and Ochang area. Among them, one bacterial strain was selected for this study based on its higher manganese oxidation, and this selected bacterial strain was identified as Aeromonas sp. MN44 through physiological-biochemical test and analysis of its 16s rRNA sequence. Aeromonas sp. MN44 was able to utilize lactose but did not utilize various carbohydrates as a sole carbon source. Aeromonas sp. MN44 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, ampicillin, tetracycline and spectinomycin, and heavy metal such as cadmium. But this strain showed a high resistance up to mg/ml unit to heavy metals such as lithium and manganese. Optimal manganese oxidation condition of Aeromonas sp. MN44 was pH 7.4 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. So, we concluded that this factor was protein. The manganese oxidizing factor produced by Aeromonas sp. MN44 was partial purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 113 kDa.

Bifidogenic Effect of Glucooligosaccharide Prepared from Glucose by Extrusion Process

  • Ahn, Jun-Bae;Hwang, Jae-Kwan;Kim, Chong-Tai;Lee, Ke-Ho;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.7 no.3
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    • pp.174-179
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    • 1997
  • In order to investigate effect of glucooligosaccharide (GOS) prepared by extrusion process as a bifidogenic factor, cultivation of Bifidobacterium sp., Bacteroides fragilis and Clostridium perfringens was done and analyzed. B. fragilis and C. perfringens were able to utilize only 16% and 11% of the oligosaccharides in GOS, respectively, whilst Bifidobacterium sp. FBD-22 could utilize 38%. Especially, many kinds of oligo saccharides in GOS were able to be utilized selectively only by Bifidobacterium sp.. In case that GOS, as a carbon source, was used in the co-cultivation by Bifidobacterium sp., B. fragilis and C. perfringens, growth of Bifidobacterium sp. was not influenced by the existence of B. fragilis and C. perfringens. Bifidobacterium sp. showed advantage on carbon source competition for GOS with B. fragilis. Acetic acid, antimicrobial agent in the intestine, was produced two times more from GOS than glucose in co-cultures of three strains. Therefore, it is suggested that GOS can be a potent bifidogenic factor which proliferates the population of Bifidobacterium sp. and may finally improve the intestinal environments of human.

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Enhancement of Lipid Production under Heterotrophic Conditions by Overexpression of an Endogenous bZIP Transcription Factor in Chlorella sp. HS2

  • Lee, Hansol;Shin, Won-Sub;Kim, Young Uk;Jeon, Seungjib;Kim, Minsik;Kang, Nam Kyu;Chang, Yong Keun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1597-1606
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    • 2020
  • Transcription factor engineering to regulate multiple genes has shown promise in the field of microalgae genetic engineering. Here, we report the first use of transcription factor engineering in Chlorella sp. HS2, thought to have potential for producing biofuels and bioproducts. We identified seven endogenous bZIP transcription factors in Chlorella sp. HS2 and named them HSbZIP1 through HSbZIP7. We overexpressed HSbZIP1, a C-type bZIP transcription factor, in Chlorella sp. HS2 with the goal of enhancing lipid production. Phenotype screening under heterotrophic conditions showed that all transformants exhibited increased fatty acid production. In particular, HSbZIP1 37 and 58 showed fatty acid methyl ester (FAME) yields of 859 and 1,052 mg/l, respectively, at day 10 of growth under heterotrophic conditions, and these yields were 74% and 113% higher, respectively, than that of WT. To elucidate the mechanism underlying the improved phenotypes, we identified candidate HSbZIP1-regulated genes via transcription factor binding site analysis. We then selected three genes involved in fatty acid synthesis and investigated mRNA expression levels of the genes by qRT-PCR. The result revealed that the possible HSbZIP1-regulated genes involved in fatty acid synthesis were upregulated in the HSbZIP1 transformants. Taken together, our results demonstrate that HSbZIP1 can be utilized to improve lipid production in Chlorella sp. HS2 under heterotrophic conditions.

Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726 (Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석)

  • Eom, Chi-Yong;Song, Seung-Eun;Park, Mi-Hwa;Kim, Young-Min
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.177-183
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    • 2007
  • Using complementation of RpoH deficient E. coli strain A7448, the rpoH gene encoding heat shock sigma factor 32 (${\sigma}^{32}$) from Methylovorus sp. strain SS1 DSM11726 was cloned and sequenced. Sequence analysis of a stretch of 1,796-bp revealed existence of an open reading frame encoding a polypeptide of 284 amino acid (32,006 dalton). Deduced amino acid sequence of the Methylovorus sp. strain SS1 RpoH showed that 59.6%, 39.1% and 51.4% identities with those of Nitrosomonas europaea (${\beta}$-proteobacteria), Agrobacterium tumefaciens ($\alpha$-proteobacteria) and E. coli (${\gamma}$-proteobacteria). The expression level of the functional ortholog of RpoH of Methylovorus sp. strain SS1 was increased transiently after heat induction, further indicating that it functions as a heat shock sigma factor.

Characterization of the porcine Nanog 5'-flanking region

  • Memon, Azra;Song, Ki-Duk;Lee, Woon Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.449-456
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    • 2018
  • Objective: Nanog, a homeodomain protein, has been investigated in humans and mice using embryonic stem cells (ESCs). Because of the limited availability of ESCs, few studies have reported the function and role of Nanog in porcine ESCs. Therefore, in this study, we investigated the location of the porcine Nanog chromosome and its basal promoter activity, which might have potential applications in development of ESCs specific marker as well as understanding its operating systems in the porcine. Methods: To characterize the porcine Nanog promoter, the 5'-flanking region of Nanog was isolated from cells of mini-pig ears. BLAST database search showed that there are two porcine Nanog genomic loci, chromosome 1 and 5, both of which contain an exon with a start codon. Deletion mutants from the 5'-flanking region of both loci were measured using the Dual-Luciferase Reporter Assay System, and a fluorescence marker, green fluorescence protein. Results: Promoter activity was detected in the sequences of chromosome 5, but not in those of chromosome 1. We identified the sequences from -99 to +194 that possessed promoter activity and contained transcription factor binding sites from deletion fragment analysis. Among the transcription factor binding sites, a Sp1 was found to play a crucial role in basal promoter activity, and point mutation of this site abolished its activity, confirming its role in promoter activity. Furthermore, gel shift analysis and chromatin immunoprecipitation analysis confirmed that Sp1 transcription factor binds to the Sp1 binding site in the porcine Nanog promoter. Taken together, these results show that Sp1 transcription factor is an essential element for porcine Nanog basal activity the same as in human and mouse. Conclusion: We showed that the porcine Nanog gene is located on porcine chromosome 5 and its basal transcriptional activity is controlled by Sp1 transcription factor.

Color Difference of Celluloytic Cultural Properties by Fungi (사상균에 의한 지류.섬유질 유물의 색변화)

  • Han, Sung-Hee;Lee, Kyu-Sik;Chung, Young-Jae
    • 보존과학연구
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    • s.17
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    • pp.48-64
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    • 1996
  • We compared the degree of color difference formed by environmental factor(temperature, relative humidity) with fungal growth in order to know how to change the color difference of cellulolutic cultural properties such as Korean papers, cotton, jute and hemp. We concluded, from the result, that the action of fungal growth on celluloytic cultural properties was more hamful than environmental factor. We considered the secretion produced by fungi as the causative agent for stained formation on cellulolytic cultural properties. Alternaria sp. colored allmaterials greyish black, Chaetomium sp. colored cotton and hemp orange, and Penicllium sp. colored cotton, jute and hemp yellowish green. But Trichoderma sp. and Aspergillus sp. didn't show a clear color against each material. It was observed that thymol(120g/$m^3$) was the most effective fungicide to prevent fungal growth.

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Characterization of the Nanog 5'-flanking Region in Bovine

  • Choi, Don-Ho;Kim, Duk-Jung;Song, Ki-Duk;Park, Hwan-Hee;Ko, Tae Hyun;Pyao, Yuliya;Chung, Ku-Min;Cha, Seok Ho;Sin, Young-Su;Kim, Nam-Hyung;Lee, Woon-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1383-1391
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    • 2016
  • Bovine embryonic stem cells have potential for use in research, such as transgenic cattle generation and the study of developmental gene regulation. The Nanog may play a critical role in maintenance of the undifferentiated state of embryonic stem cells in the bovine, as in murine and human. Nevertheless, efforts to study the bovine Nanog for pluripotency-maintaining factors have been insufficient. In this study, in order to understand the mechanisms of transcriptional regulation of the bovine Nanog, the 5'-flanking region of the Nanog was isolated from ear cells of Hanwoo. Results of transient transfection using a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the -134 to -19 region contained the positive regulatory sequences for the transcription of the bovine Nanog. Results from mutagenesis studies demonstrated that the Sp1-binding site that is located in the proximal promoter region plays an important role in transcriptional activity of the bovine Nanog promoter. The electrophoretic mobility shift assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. In addition, significant inhibition of Nanog promoter activity by the Sp1 mutant was observed in murine embryonic stem cells. Furthermore, chromatin-immunoprecipitation assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. These results suggest that Sp1 is an essential regulatory factor for bovine Nanog transcriptional activity.

AUA as a Translation Initiation Site In Vitro for the Human Transcription Factor Sp3

  • Hernandez, Eric Moore;Johnson, Anna;Notario, Vicente;Chen, Andrew;Richert, John R.
    • BMB Reports
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    • v.35 no.3
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    • pp.273-282
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    • 2002
  • Sp3 is a bifunctional transcription factor that has been reported to stimulate or repress the transcription of numerous genes. Although the size of Sp3 mRNA is 4.0kb, the size of the known Sp3 cDNA sequence is 3.6kb. Thus, Sp3 functional studies have been performed with an artificially introduced start codon, and thus an amino-terminus that differs from the wild-type. Ideally, full-length cDNA expression vectors with the appropriate start codon should be utilized for these studies. Using 5'rapid amplification of cDNA ends, a full-length Sp3 cDNA clone was generated and the sequence verified in nine cell lines. No AUG initiation codon was present. However, stop codons were present in all three frames 5' to the known coding sequence. In vitro translation of this full-length cDNA clone produced the expected three isoforms-one at 100 kDa and two in the mid 60 kDa range. Electrophoretic mobility shift assays showed that the protein products had the ability to bind to the Sp1/3 consensus sequence. In vitro studies, using our Sp3 clone and site directed mutagenesis, identified the translation initiation site for the larger isoform as AUA. AUA has not been previously described as an endogenous initiation codon in eukaryotes.