• BMB Reports
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• v.36 no.1
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• pp.35-42
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• 2003

Heterocyclic amines (HCAs) are potent mutagens generated during the cooking of meat and fish, and several of these compounds produce tumors in conventional experimental animals. During the past 5 years or so, HCAs have been tested in a number of novel in vivo murine models, including the following: lacZ, lacI, cII, c-myc/lacZ, rpsL, and $gpt{\Delta}$ transgenics, $XPA^{-/-}$, $XPC^{-/-}$, $Msh2^{+/-}$, $Msh2^{-/-}$ and $p53^{+/-}$ knock-outs, Apc mutant mice ($Apc^{{\Delta}716}$, $Apc^{1638N}$, $Apc^{min}$), and $A33^{{\Delta}N{\beta}-cat}$ knock-in mice. Several of these models have provided insights into the mutation spectra induced in vivo by HCAs in target and non-target organs for tumorigenesis, as well as demonstrating enhanced susceptibility to HCA-induced tumors and preneoplastic lesions. This review describes several of the more recent reports in which novel animal models were used to examine HCA-induced mutagenesis and carcinogenesis in vivo, including a number of studies which assessed the inhibitory activities of chemopreventive agents such as 1,2-dithiole-3-thione, conjugated linoleic acids, tea, curcumin, chlorophyllin-chitosan, and sulindac.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.872-880
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• 2003

Three vectors, pSG1, 2, and 3, which facilitate in vivo expression technology (IVET) in Gram-negative bacteria, were developed. Vectors pSG1and 2 are derivatives of ColE1, and pSG3 is a derivative of an R6K replicon. These vectors contain oriT sites that allow mobilization when the RK2 Tra functions are provided in trans. These vectors contain promoterless lacZ (pl-lacZ) and promoterless aph (pl-aph) transcriptionally fused together, which allow qualitative and quantitative measurements of the expression of genes in the genome of bacterial cells. pSG1 and 3 contain gentamicin-resistance genes, and pSG2 carries a streptomycin-/spectinomycin-resistance gene, allowing for selection of recombinants generated by a single crossover between a library fragment cloned into a pSG vector and the identical region in the genome of a bacterial species from which the library fragment originated. These vectors were successfully applied to the generation of random fusions at high rates in the genomes of four representative Gram-negative bacteria. In addition, the expression level of ${\beta}-galactosidase$ and the degree of resistance to kanamycin in cells with fusions generated by these vectors were found to be linearly correlated, proving that these vectors can be used for IVET.

• Environmental Engineering Research
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• v.14 no.1
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• pp.63-67
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• 2009

Escherichia coli K-12 (E. coli K-12) is a representative indicator globally used for distinguishing and monitoring dynamic fates of pathogenic microorganisms in the environment. This study investigated how to most critically quantify lacZ ($\beta$-galactosidase) gene in E. coli K-12 by two different real-time polymerase chain reaction (real-time PCR) in association with three different DNA extraction practices. Three DNA extractions, i.e., sodium dodecyl sulfate (SDS)/proteinase K, magnetic beads and guanidium thiocyanate (GTC)/silica matrix were each compared for extracting total genomic DNA from E. coli K-12. Among them, GTC/silica matrix and magnetic beads beating similarly worked out to have the highest (22-23 ng/${\mu}L$) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/${\mu}L$) concentration of DNA retrieved. There were no significant differences in the quantification of the copy numbers of lacZ gene between SYBR Green I qPCR and QProbe-qPCR. However, SYBR Green I qPCR obtained somewhat higher copy number as $1{\times}10^8$ copies. It was decided that GTC/silica matrix extraction or magnetic beads beating in combination with SYBR Green I qPCR can be preferably applied for more effectively quantifying specific gene from a pure culture of microorganism.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.778-782
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• 2007

This study assessed the effects of the antimicrobial substances sulforaphane, grapefruit seed extracts (GSE), and reuterin on the expression of Salmonella HilA and InvF virulence gene using a LacZY assay (${\beta}$-galactosidase assay) with hilA:lacZY and invF:lacZY fusion strains of Salmonella typhimurium SL1344. Salmonella was grown for 8 hr at $37^{\circ}C$ in the presence of diluted antimicrobial substances ($2\;{\mu}g/mL$ sulforaphane, $20\{\mu}g/mL$ GSE, and 0.26 mM reuterin) at concentrations that did not inhibit the cellular growth of Salmonella. Sulforaphane inhibited the expression of HilA and InvF by 50-90 and 20-80%, respectively. GSE also inhibited the expression of both genes, but to a lesser degree. Among the 3 antimicrobial substances, reuterin showed the least inhibition, which was abolished after 3-4 hr. None of the antimicrobial substances inhibited the ${\beta}$-galactosidase enzyme activity of S. typhimurium. The assay used in this study represents a very sensitive method for screening bioactive substances that inhibit the expression of virulence genes in Salmonella.

• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.373-378
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• 1998

This study was performed to investigate antimicrobial effects and their actions of the ethanol extracts of 12 spices against Lactobacillus plantarum and Leuconostoc mesenteroides, which are related to Kimchi fermentation. The cardamon, thyme, and cumin ethanol extracts among 12 spices showed antimicrobial activities against Lac. plantarum, not against Leu. mesenteroides through paper disc method. The growth of Lac. plantarum was inhibited in MRS broth containing each extracts of cardamon (>4%), thyme (>4%) and cumin (>1%). SDS-PAGE and transmission electron micrographs showed that the cell walls and membranes were disrupted and the cytoplasmic components were leaked in strains treated with ethanol extracts.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.63-68
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• 2000

GroEL is well known as a molecular chaperone. In order to determine the dynamic stress response of the Escherichia coli groE promoter, a groE-lacZ operon fusion in the chromosome was constructed. Stress leading to ${\sigma}^{32}$ synthesis induces transcription from E. coli groE promoter, since the promoter is ${\sigma}^{32}-regulated$. When the strain was stressed with ethanol, phenol, and sodium chloride, clear inductions of ${\beta}-galactosidase$ were observed. Two types of simultaneous stresses of sodium chloride and phenol induced the enze much more than either of the two alone, suggesting that stress was an additive. The combined stress resulted in the highest induction of the enzyme in this system. The groE-lacZ fusion strain developed in this study can conveniently be used to detect other harmful pollutants in the environment. Stress treatment of cells containing recombinant proteins, which need GroEl, by ethanol, phenol, or sodium chloride, might have a tendency to increase their biological activities.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.144-150
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• 1997

After DNA damage, umuDC is the only SOS operon that must be induced to promote SOS mutagenesis in Escherichia coli. The recombinant plasmid pBC401 and pBC402 were constructed to fuse the lac structural genes with promoter region of umuDC operon to induce the expression of lacZ gene by DNA damage. We transformed the plasmid pBC401 and pBC402 into E. coli MC1061, lacZ deleted strain and determined the activity of $\beta$-galactosidase for various mutagen; UV, mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroqunoline-1-oxide (NQO), ethyl methanesulfonate (EMS). The $\beta$-galactosidase activities of PBC401 and pBC402 for UV, MMC, and NQO were increased in proportion to expression time until 3 hours thereafter, the activities were constant or slightly decreased. The activities for MNNG and EMS were not so high as for UV, MMC, and NQO. When MNNG and EMS were treated, $\beta$-galactosidase activity of pBC402 was slightly lower than pBC401 but when UV, MMC, and NQO were treated in pBC402, $\beta$-galactosidase activity was slightly higher than in pBC401. Therefore, the pBC402 was better than the pBC401 in terms of sensitivity for frameshift mutagen. We suggest that the plasmid pBC401 and pBC402 are easy to detect mutagens which cause frameshift mutation rather than point mutation.

• The Journal of Engineering Research
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• v.8 no.1
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• pp.31-50
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• 2006

Shrinkage may cause cracking in concrete. In practice such cracking must be considered in most concrete applications because, under normal conditions, drying of the concrete is unavoidable, and when drying takes place shrinkage occurs. Cracked concrete is an inferior concrete because it is weaker, more permeable, and more susceptible to chemical attack. The development of the strength of LAC with aging depends on a few factors such as type of the cement, W/C ratio, curing conditions and periods. The higher the strength of LAC, the lower the possibility of shrinkage cracking. Hence, the strength of LAC in the hypocaust system depends to a large extent on the effect of cracking decrease of the antifoaming rate to drying shrinkage in cement.

• Genomics & Informatics
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• v.1 no.2
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• pp.113-118
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• 2003

Among a number of antigens characterized in M leprae, an etiological agent of Leprosy, the 18 kDa antigen, is unique to M leprae. We have previously determined a sequence specific element in the 18 kDa gene of M leprae, which confers transcriptional repression. In this report, we have examined if the element could be applied to genes other than the 18 kDa gene of M leprae. To identify the roles of the regulatory sequence in heterologous promoter, we have constructed pB3 vector series, which contains BCG hsp65 promoter and the M leprae 18 kDa transcription repression responsive element in tandem using LacZ gene as a reporter gene. Cloning of hsp65 promoters of M bovis BCG or M smegmatis in front of LacZ gene resulted in normal $\beta$­galactosidase activity as expected. However, when the sequence element was placed between the promoter and the LacZ gene, $\beta$-galactosidase activity was reduced 10-fold less. Also we have examined with pB3(-) vector, that harbors the transcription repression responsive element in a reversed orientation, the $\beta$-galactosidase activity was found to be similar to pB3(+) vector. Thus, these results further confirm that M leprae 18 kDa transcription repression responsive element could regulate BCG hsp65 heterologous promoter and that the element could act as an operator for the transcription of mycobacteria.

• Journal of the Korean Wood Science and Technology
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• v.27 no.4
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• pp.72-79
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• 1999

The ability of several wood rotting fungi for decolorization of two anthracene derivatives, Carminic acid (CA) and Remazol brilliant blue R (RBBR), and hardwood KP bleaching liquor (BL) as well as laccase activities in these fungi were studied. The enzyme activity appeared exclusively in fungi destaining RBBR and CA, but in the case of BL, such relationship was not observed. The laccase enzyme was released into the decolorization media and its inducible (but not constitutive) forms shown destaining activity. The purified inducible forms of Kuehneromyces mutabilis and Pleurotus ostreatus laccase destained CA. Thus the possible differentiation between specificity of particular LAC forms was confirmed. In addition the nitrogen starvation induced both laccase and CA destaining activities, but the increase was higher for decolorization of CA than LAC activity. Probably LAC would be only partly responsible for decolorization of this dye. This results suggested that purified LACs decolorize CA, however its destaining activities were considerably lower than the activities on syringaldazine.