• Title/Summary/Keyword: sM gene

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Induction of Microsomal Epoxide Hydrolase, rGSTA2, rGSTA3/5, and rGSTM1 by Disulfiram, but not by Diethyldithiocarbamate, a Reduced Form of Disulfiram

  • Kim, Sang-Geon;Kim, Hye-Jung
    • Toxicological Research
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    • v.13 no.4
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    • pp.339-347
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    • 1997
  • Disulfiram (DSF) and diethyldithiocarbamate (DDC), a reduced form of DSF, protect the liver against toxicant-induced injury through inhibition of cytochrome P450 2E1. The effect of DSF and DDC on the levels of major hepatic microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) expression was comparatively studied, given the view that these enzymes are involved in terminal detoxification events for high energy intermediates of xenobiotics. Treatment of rats with a single dose of DSF (20-200 mg/kg, po) resulted in 2- to 15-fold increases in the mEH mRNA level at 24 hr with the ED$_{50}$ value being noted as 60 mg/kg. The mEH mRNA level was elevated ~15-fold at 24 hr after treatment at the dose of 100 mg/kg, whereas the hepatic mRNA level was rather decreased from the maximum at the dose of 200 mg/kg, indicating that DSF might cause cytotoxicity at the dose. In contrast to the effect of DSF, DDC only minimally elevated the mEH mRNA level at the doses employed. DSF moderately increased the major GST mRNA levels in the liver as a function of dose, resulting in rGSTA2, rGSTA3/5 or rGSTM1 mRNA levels being elevated 3- to 4-fold at 24 hr post-treatment, whereas the rGSTM2 mRNA level was not altered. DDC, however, failed to stimulate the mRNA levels for major GST subunits, indicating that the reduced form of DSF was ineffective in stimulating the GST the expression. The effect of other organosulfides including aldrithiol, 2, 2'-dithiobis(benzothiazole) (DTB), tetramethylthiouram disulfide (TMTD) and allyl disulfide (ADS) on the hepatic mEH and GST mRNA expression was assessed in rats in order to further confirm the increase in the gene expression by other disulfides. Treatment of rats with aldrithiol (100 mg/kg, po) resulted in a 16-fold increase in the mEH mRNA level at 24 hr post-treatment. DTB, TMTD and ADS also caused 5-, 9- and 12-fold increases in the rnRNA level, respectively, as compared to control. Thus, all of the disulfides examined were active in stimulating the mEH gene in the liver. The organosulfides significantly increased the rGSTA2, rGSTA3, rGSTA5 and rGSTM1 mRNA levels at 24 hr after administration. In particular, aldrithiol was very efficient in stimulating the rGSTA and rGSTM genes among the disulfides examined. These results provide evidence that DSF and other sulfides effectively stimulate the mEH and major GST gene expression at early times in the liver and that DDC, a reduced form of DSF, was ineffective in stimulating the expression of the genes, supporting the conclusion that reduced form(s) of organosulfur compound(s) might be less effective in inducing the mEH and GST genes through the antioxidant responsive element(s).

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Assessment of Bile Salt Effects on S-Layer Production, slp Gene Expression and, Some Physicochemical Properties of Lactobacillus acidophilus ATCC 4356

  • Khaleghi, M.;Kermanshahi, R. Kasra;Yaghoobi, M.M.;Zarkesh-Esfahani, S.H.;Baghizadeh, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.749-756
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    • 2010
  • In many conditions, bacterial surface properties are changed as a result of variation in the growth medium and conditions. This study examined the influence of bile salt concentrations (0-0.1%) on colony morphotype, hydrophobicity, $H_2O_2$ concentration, S-layer protein production, and slpA gene expression in Lactobacillus acidophilus ATCC 4356. It was observed that two types of colonies (R and S) were in the control group and the stress condition. When the bile level increased in the medium, the amount of S type was more than the R type. A stepwise increment in the bile concentration resulted in a stepwise decline in the maximum growth rate. The results showed that hydrophobicity was increased in 0.01%-0.02% bile, but it was decreased in 0.1% bile. Treatment by bile (0.01%-0.1%) profoundly decreased $H_2O_2$ formation. S-Layer protein and slpA gene expression were also altered by the stress condition. S-Protein expression was increased in the stress condition. The slpA gene expression increased in 0.01%-0.05% bile and it decreased in 0.1% bile. However, we found that different bile salt concentrations influenced the morphology and some surface properties of L. acidophilus ATCC 4356. These changes were very different in the 0.1% bile. It appears that the bacteria respond abruptly to 0.1% bile.

Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.77-82
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    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

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Molecular Sexing Using SRY and ZF Genes in Pigs (돼지 SRY와 ZF 유전자를 이용한 성판별 기법)

  • Cho, I.C.;Kang, S.Y.;Lee, S.S.;Choi, Y.L.;Ko, M.S.;Oh, M.Y.;Han, Sang-Hyun
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.317-324
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    • 2005
  • A method for sex determination of pigs was examined using polymerase chain reaction(PCR). Sex determining region Y(SRY) gene encoded on Y chromosome plays a key role for primary male development. Zinc finger X-Y(ZFX-ZFY) gene, one of the X-V homology gene group was found on the X and Y chromosomes, respectively, We tested for molecular sexing by amplification patterns of SRY and ZF genes. Genomic DNAs from various resources including porcine hairs and semen collected from domestic pig breeds and native pigs was used for PCR assay of each gene. The amplified products for porcine SRY gene were yielded only in males but not in females. On the other hand, two differential patterns were observed in amplification of ZF gene reflecting the chromosomal dimorphism by a length polymorphism between X and Y chromosomes. Of both, a common band was detected in all individuals tested so that this band might be amplified from ZFX gene as a PCR template, but another is specific for males indicated that from ZFY. The result of PCR assay provides identical information to that from investigation of phenotypic genders of the pigs tested. We suggest that this PCR strategy to determine porcine sexes using comparison of the amplification patterns of the SRY gene specific for Y chromosome and the dimorphic ZF gene between X and Y chromosomes may be a rapid and precise method for discrimination of two sexes and applied to DNA analysis of small samples such as embryonic blastomere, semen, and hairs.

Schizosaccharomyces pombe nup97, which Genetically Interacts with mex67, is Essential for Growth and Involved in mRNA Export

  • Cho, Hyun-Jin;Hwang, Duk-Kyung;Jung, Sun-Im;Yoon, Jin-Ho
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.344-349
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    • 2007
  • We have isolated previously three synthetic lethal mutants in Schizosaccharomyces pombe, which genetically interact with mex67, in order to identify the genes involved in mRNA export. A novel nup97 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex3. The nup97 gene contains one intron and encodes an 851 amino-acid protein that is similar to nucleoporins, Nppl06p in S. pombe and Nic96p in Saccharomyces cerevisiae. The nup97 gene is essential for vegetative growth, and nup97 null mutant harboring pREP41X-Nup97 showed $poly(A)^+$ RNA export defect when expression of nup97 is repressed in the presence of thiamine. These results suggest that nup97 is involved in mRNA export from the nucleus to cytoplasm.

Lack of Association of Glutathione S-transferase M3 Gene Polymorphism with the Susceptibility of Lung Cancer

  • Feng, Xu;Dong, Chun-Qiang;Shi, Jun-Jie;Zhou, Hua-Fu;He, Wei;Zheng, Bao-Shi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4465-4468
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    • 2012
  • Objective: The conclusions of published reports on the relationship between the glutathione S-transferase M3 (GSTM3) A/B gene polymorphism and the risk of lung cancer are still debated. This meta-analysis was performed to evaluate the association between GSTM3 and the risk of lung cancer. Methods: Association investigations were identified from PubMed, Embase, and Cochrane Library, and eligible studies were included and synthesized using a meta-analysis method. Results: Eight reports were included into this meta-analysis for the association of GSTM3 A/B gene polymorphism and lung cancer susceptibility, covering 1,854 patients with lung cancer and 1,926 controls. No association between the GSTM3 A/B gene polymorphism and lung cancer was found in this meta-analysis (B allele: OR = 1.25, 95% CI: 0.89-1.76, P = 0.20; BB genotype: OR = 1.53, 95% CI: 0.71-3.32, P = 0.28; AA genotype: OR = 0.85, 95% CI: 0.59-1.23, P = 0.39). Conclusions: The GSTM3 A/B gene polymorphism is not associated with lung cancer susceptibility. However, more studies on the relationship between GSTM3 A/B gene polymorphism and the risk of lung cancer should be performed in the future.

Cloning and Transctiption of Excherichia coli Cell Division Gene, sep (E. coli 세포분열 유전자 sep의 Cloning 및 Transcription에 관한 연구)

  • ;Walker, James R.
    • Korean Journal of Microbiology
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    • v.22 no.4
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    • pp.235-242
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    • 1984
  • Sep gene, which is one of the cell division genes coding for penicillin binding protein 3 was subcloned from ${\lambda}607sep^{+2}$ to plasmid pBR322. which has a strong promotor such as lac UV5(lacP). It was confirmed that the sep gene cloned to pLJ3 was in the proper orientation for expressionfrom lactose promotor. To analyze the expression efficiency of sep gene within the plasmids newly constructed, sep mRNA was assated by using ${\lambda$\mid$\;607sep^{+2}$ DNA as a probe. Sep mRNA level was increased 25 times in the cells carrying sep gene cloned to pBR322 compared to E. coli C600 which has wild type sep gene within the chromosome instead of plasmed. Furthermore, the cells carrying sep gene cloned to pLJ3 derected the synthesis of about 50 times as much sep mRNA as did cells carrying sep gene cloned to pBR 322, representing that the sep gene was successfully cloned to pLJ3.

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Effects of spTho1 Deletion and Over-Expression on mRNA Export in Fission Yeast (분열효모에서 spTho1 유전자의 결실과 과발현이 생장 및 mRNA Export에 미치는 영향)

  • Cho, Ye-Seul;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.401-404
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    • 2010
  • Tho1 is a RNA-binding protein that assembles co-transcriptionally onto the nascent mRNA and is thought to be involved in mRNP biogenesis and mature mRNA export to cytoplasm in budding yeast. In fission yeast Schizosaccharomyces pombe, a homologue of THO1 (spTho1) was identified based on sequence alignment. A deletion mutant in a diploid strain was constructed by replacing one of spTho1-coding region with an ura4+ gene using one-step gene disruption method. Tetrad analysis showed that the spTho1 was not essential for growth. The spTho1 mutant did not show any defects of bulk mRNA export. However, over-expression of spTho1 from strong nmt1 promoter caused the growth defects and accumulation of poly(A)$^+$ RNA in the nucleus. These results suggest that spTho1 is involved in mRNA export from the nucleus to cytoplasm though it is not essential.