• Title/Summary/Keyword: S1 gene

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Nucleotide sequence analysis of the 5S ribosomal RNA gene of the mushroom tricholoma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.136-141
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    • 1995
  • From a cluster of structural rRNA genes which has previsouly been cloned (Hwang and Kim, in submission; J. Microbiol. Biotechnol.), a 1.0-kb Eco RI fragment of DNA which shows significant homology to the 25S and rRNA s of Tricholoma matsutake was used for sequence analysis. Nucleotide sequence was bidirectionally determined using delection series of the DNA fragment. Comparing the resultant 1016-base sequence with sequences in the database, both the 3'end of 25S-rRNA gene and 5S rRNA gene were searched. The 5S rRNA gene is 118-bp in length and is located 158-bp downstream of 3'end of the 25S rRNA gene. IGSI and IGS2 (partial) sequences are also contained in the fragment. Multiple alignment of the 5S rRNA sequences was carried out with 5S rRNA sequences from some members of the subdivision Basidiomycotina obtained from the database. Polygenetic analysis with distance matrix established by Kimura's 2-parameter method and phylogenetic tree by UPGMA method proposed that T. matsutake is closely related to efibulobasidium allbescens. Secondary structure of 5S rRNA was also hypothesized to show similar topology with its generally accepted eukaryotic counterpart.

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Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

  • Chung, Injae
    • Toxicological Research
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    • v.16 no.2
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    • pp.95-100
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    • 2000
  • In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

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Lack of Assocation of Glutathione S-transferase T1 Gene Null and Susceptibility to Lung Cancer in China: a Meta-analysis

  • Liu, Hong-Zhou;Peng, Jie;Zheng, Fang;Wang, Chun-Hong;Han, Ming-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7215-7219
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    • 2013
  • Background: Variation in metabolic genes is regarded as an important factor in processes leading to cancer. However, the effect of GSTT1 null genotype is divergent in the form of lung cancer. Methods: Studies were conducted at different research databases from 1990 to 2013 and the total odds ratio (OR) and 95% confidence interval (CI) were calculated for lung cancer. Review Manager 5.2 and STATE 12 are employed. Results: Total OR value is calculated from 17 articles with 2,118 cases and 2,915 controls. We discovered no significant increase in lung cancer risk among subjects carrying GSTT1 null genotype [OR = 1.15; 95% CI 0.97-1.36] in this meta-analysis. Conclusion: The GSTT1 deletion polymorphism does not have a significant effect on the susceptibility to lung cancer overall in China.

Identification of a Novel Fusion Gene (HLA-E and HLA-B) by RNA-seq Analysis in Esophageal Squamous Cell Carcinoma

  • Jiang, Yu-Zhang;Li, Qian-Hui;Zhao, Jian-Qiang;Lv, Jun-Ji
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2309-2312
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    • 2014
  • Esophageal squamous cell carcinoma (ESCC) is the most common histologic subtype of esophageal cancer and is characterized by a poor prognosis. Determining gene changes in ESCCs should improve understanding of putative risk factors and provide potential targets for therapy. We sequenced about 55 million pair-end reads from a pair of adjacent normal and ESCC samples to identify the gene expression level and gene fusion. Sanger sequencing was used to verify the result. About 17 thousand genes were expressed in the tissues, of which approximately 2400 demonstrated significant differences between tumor and adjacent non tumor tissue. GO and KEGG pathway analysis revealed that many of these genes were associated with cellular adherence and movement, simulation responses and immune responses. Notably we identified and validated one fusion gene, HLA-E and HLA-B, located 1 MB apart. We also identified thousands of remarkably expressed transcripts. In conclusion, a novel fusion gene HLA-E and HLA-B was identified in ESCC via whole transcriptome sequencing, which would be a biomarker for ESCC diagnosis and target for therapy, shedding new light for better understanding of ESCC tumorigenesis.

Comparison of scanning electron microscopic structures and nucleotide sequences variation of ITS1, 5.8S ribosomal RNA gene and ITS2 region in three Peruvian entomopathogenic fungal isolates (3종의 페루산 entomopathogenic fungi의 전자현미경적 구조와 ITS1, 5.8S ribosomal RNA gene, ITS2의 염기서열 다양성)

  • Han, Sang-Hoon;Nam, Sunghee;Lee, Heui-Sam;Yeo, Joo-Hong
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.137-141
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    • 2013
  • In this study, nucleotide sequence structures of intergenic transcribed spacer (ITS) 1, complete 5.8S ribosomal RNA gene and ITS 2 region were analyzed to identify three Peruvian entomopathogenic fungal isolates. The isolates had highly conserved sequence region in 5.8S rRNA gene and unique sequences in ITS 1 and 2 region among them. 5.8S rRNA gene regions were highly conserved and showed high homoloies among tested isolates. In contrast, ITS region showed species-specific sequence region, resulting in inter-genus differencies. Scanning electron microscopic images of these isolates supported the result of ITS-based identification. From these result, Peruvian entomopathogenic fungal isolate J270, J278, were identified as Beauveria bassiana and J271 was identified as Lecanicillium attenuatum.

The biochemical and molecular characteristics of Streptococcus equi subsp. zooepidemicus isolated from the genital tract of Thoroughbred mares in Korea

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Cho, Gil-Jae
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.201-208
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    • 2011
  • Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a pathogen of a variety of infections in horse. We studied biochemical and molecular characteristics of S. zooepidemicus isolated from the genital tract of Thoroughbred mares in Korea. Seventy-nine isolates were identified as S. zooepidemicus by biochemical and PCR method from 374 horses. The biochemical characteristics of S. zooepidemicus isolates were positive reaction of lactose and sorbitol. However, S. zooepidemicus isoltes were negative reaction of inulin, mannitol, raffinose, trehalose, aesculin hydrolysis, growth in 6.5% NaCl and variable reaction of maltose. Epidemiological investigations of S. zooepidemicus isolates were performed by fragment analysis of SzP (S. zooepidemicus protective protein) gene, CNE (collagen binding protein) gene and ISR (16s rRNA intergenic spacer region) gene using ABI Prism $3,130{\times}1$ Genetic Analyzer System. All isolates were shown single amplification size of 906 bp in CNE gene, but SzP and ISR gene were shown variable patterns of fragment size. The characteristics of S. zooepidemicus investigated in this study will be very useful for the prevention of infection and the studies of epidemiologic characteristics of S. zooepidemicus, causing the severe economic losses due to reproductive failures.

Characterization of the pcbE Gene Encoding 2-Hydroxypenta-2,4-Dienoate Hydratase in Pseudomonas sp. DJ-12

  • Lim, Jong-Chul;Lee, Jeongrai;Jang, Jeong-Duk;Lim, Jai-Yun;Min, Kyung-Rak;Kim, Chi-Kyung;Kim, Young-Soo
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.187-195
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    • 2000
  • Nucleotide sequence extending 2,3-dihydroxybiphenyl 1,2-dioxygenase gene (pcbC) and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase gene (pcbD) of Pseudomonas sp. DJ-12 was previously analyzed and the two genes were present in the order of pcbD-pcbC preceded by a promoter from Pseudomonas sp. DJ-12. In this study, a 3.8-kb nucleotide sequence located downstream of the pcbC gene was analyzed to have three open reading frames (ORFs) that are designated as orf1, pcbE and orf2 genes. All of the ORFs were preceded by each ribosome-binding sequence of 5-GGAXA-3 (X=G or A). However, no promoter-like sequence and transcription terminator sequence were found in the analyzed region, downstream of pcbC gene. Therefore, the gene cluster appeared to be present in the order of pcbD-pcbC-orf1-pcbE-orf2 as an operon, which is unique organization characterized so far in biphenyl- and PCB-degrading bacteria. The orf1 gene was composed of 1,224 base pairs which can encode a polypeptide of molecular weight 44,950 containing 405 amino acid residues. A deduced amino acid sequence of the orf1 gene product exhibited 21-33% identity with those of indole dioxygenase and phenol hydroxylase components. The pcbE gene was composed of 783 base pairs encoding 2-hydroxypenta-2,4-dienoate hydratase involved in the 4-chlorobiphenyl catabolism. The orf2 gene was composed of 1,017 base pairs encoding a polypeptide of molecular weight 37,378 containing 338 amino acid residues. A deduced amino acid sequence of the orf2 gene product exhibited 31% identity with that of a nitrilotriacetate monooxygenase component.

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Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector I. Expression of cloned amylase gene in Escherichia coli (YRp 7 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 서정훈;김영호;전도연;홍순덕;조윤래
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.161-168
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    • 1986
  • A 1.95Kb Sau3Al fragment coding for $\alpha$-amylase from Bacillus amyloliquefaciens was isolated by the shotgun method using Escherichia coli as a host. The genome of Bacillus amyloliquefaciens was partially digested with the restriction endonuclease Sau3Al and joined to plasmid YRp7 cleaved with the restriction endonuclease BamHI. The $\alpha$-amylase gene present in a 1.95Kb insert was stably maintained and expressed in Escherichia coli. The amount of $\alpha$-amylase activity produced by Escherichia coli containing the hybrid plasmid pEA24 was about 65% of the activity produced by the donor Bacillus amyloliquefaciens strain. The properties of $\alpha$-amylase produced by Escherichia coli were very similar to those produced by Bacillus amyloliquefaciens as based on optimum temperature, pH, and effect of CaCl$_2$ concentration. About 70% of the $\alpha$-amylase produced by Escherichia coli was localized in the periplasmic space, whereas the remaining enzyme was localized in the inner part of the cell.

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Diagnosis of Salmonella dubin in Korean Native Calves using PCR and Nucleotide Sequences of rfb5 Gene (송아지에 감염된 Salmonella dublin의 PCR 진단과 rfbS 항원단백 유전자의 염기서열분석)

  • 김철민;이영준;박명규;최경성;김민석
    • Journal of Veterinary Clinics
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    • v.17 no.2
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    • pp.464-469
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    • 2000
  • An epizootic of calf diarrhea occurred in a Korean native cattle farm located in Chonbuk province. The calves that had either bloody or watery diarrhea were 1 to 30 days old. Some of these animals died during the acute course of the disease. Five calves with predominant clinical signs were examined in more detail. Hematological and serum chemical findings were suggestive of dehydration and nutritional insufficiency. Fecal material from the calve was cultured on/in brilliant green agar (BGA), xylose-lysine deoxycholate (XLD) medium, MacConkey agar, eosin methylene blue (EMB) agar and triple sugar iorn (TSI) A bacteria was isolated. which was subsequently identificed as belonging to Salmonella spp. To differentiate Salmoenlla serotype, rfbs gene of S. dublin was amli- find (720 bp) by multiplex (PCR). The rfbS gene sequences of S, dublin ficld isolate(SDC-1) was com- pared with that off S. dublin(S-37) S, dublin(Ahn et al, 1996), S enteritidis(Ahn et al 1996)and S. typhi (Generbak accession No M29682). The identities of nucleotide sequences were 100%. 99.6%, 99.6%, 97.5% respectively.

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Promoter Structure and Transcriptional Activity of Human Complement Receptor Type I (CR1) Gene

  • Kim, Jae-Hyun;Lee, Young-Ju;Nam, Ju-Ryoung;Shim, Hee-Bo;Choe, Soo-Young
    • Animal cells and systems
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    • v.7 no.1
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    • pp.63-68
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    • 2003
  • Until recently, interest in human complement receptor type I (CR1) has focused on immune complex processing, which contributed to our understanding of regulatory mechanism of complement activation. However, the promoter structure and transcriptional regulation of human CR1 gene has not been clear. To study the unique regulation of human CR1 gene expression, we assessed promoter activity of the $5^1$-flanking region of human CR1 gene using transient transfection and gel mobility shift assays. In this study we demonstrated that NF-Y binds to the inverted CCAAT element and that the functional interaction with protein(s) which bind to the GC-rich motif may be necessary for optimal transcription of human CR1 gene. We also show that sequence elements which located at-95/58 and +45/+50 are important for optimal transcription of CR1 gene.