• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.191-196
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• 2004

In this study, to construct more effective retrovirus system, we compared four internal promoters (RSV: Rous sarcoma virus, UbC: Ubiquitin C, β-actin, CMV: human cytomegalovirus) in the retrovirus vector by measuring GFP (green fluorescent protein) expression. The effect of WPRE (woodchuck hepatitis virus posttrans-criptional regulatory element) sequence on transgene expression was also investigated. Experiments were conducted with cells derived from three different species (human, pig and chicken) and evaluated the activity of each promoter and the effect of WPRE sequence by fluorometry and Western blotting. In all cells tested, RSV and CMV promoters were superior to UbC and β-actin promoters, and RSV promoter was the best one in chicken cells. The boosting effect of WPRE sequence on GFP expression was evident in human and porcine cells but not in chicken cells.

• Journal of Microbiology and Biotechnology
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• v.3 no.3
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• pp.146-155
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• 1993

A DNA fragment from an alkali-tolerent Bacillus sp., conferring strong promoter activity, was subcloned into the promoter probe plasmid pPL703 and the nucleotide sequence of this promoter region was determined. The sequence analysis suggested that this highly efficient promoter region containing the complex clustered promoters comprised three kinds of promoters (P1, P2 and P3), which are transcribed by $\sigma^B (formerly \sigma^{37}), \sigma^E(formerly \sigma^{29}) and \sigma^A (formerly \sigma^{43})$ RNA polymerase holoenzymes which play major rules at the onset of endospore formation, during sporulation and at the vegetative phase of growth, respectively. S1 nuclease mapping experiments showed that all three promoters had staggered transcription initiation points. The results of chloramphenicol acetyltransferase assay after the subcloning experiments also indicated that the expression of these clustered promoters was correlated with the programs of growth and endospore development. Promoter P1, P2 and P3 were preceded by 75% AT, 79% AT and 81% AT regions, respectively, and a partial deletion of AT-rich region prevented transcription from promoter P1 in vivo. Two sets of 5 -AGTGTT-3 sequences and inverted repeat sequences located around the promoter P1 were speculated as the possible cis acting sites for the catabolite repression in B. subtilis. In vivo transcripts from these sequence regions may be able to form a secondary structure, however, the possibility that a regulatory protein induced by the excess amount of glucose could be bound to such a domain for crucial action remains to be determined.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.461-465
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• 2006

We investigated the protein productivity of the promoters for genes showing prolonged induction upon cold shock in Escherichia coli. Six low temperature inducible genes (frdA, glpB, hypB, katG, nupG, ompT) were selected based on the previously reported cDNA microarray based global transcription profiling of Escherichia coli Kl2 in response to cold shock. Their promoter regions were isolated from the genomic DNA of E. coli JM109 and expression levels induced by the promoters were examined by using green fluorescence protein (GFP) as a reporter at $15^{\circ}C$ and $37^{\circ}C$. Among the six promoters, the promoter for nupG showed the highest and prolonged expression at both temperatures and the cold inducibility of nupG promoter was not observed.

• Journal of Food Hygiene and Safety
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• v.15 no.1
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• pp.51-54
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• 2000

In the 5'-flanking region of the D-AAT, AspAT and AlaDH gene, I found three or two pairs of sequences(designated as Pl, P2, P3) which show significant similarity to the E.coli consensus sequences of -35 and -10 for promoters. The spacing between -35 and -10 is 16 to 18bp in all the three putative promoters Pl, P2 and P3 which is in good agreement with the preferred spacer length in E.coli and in B.subtilis. Therefore, the putative promoters may also function to increase the efficiency of transcriptional initiation. The most stable, double-helical“Shine-Dalgarno”pairing is formed with a free energy change(ΔG) of -13.0 kcal/mol, -9.6 kcal/mol, -15.8 kcal/mol.

• Microbiology and Biotechnology Letters
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• v.16 no.5
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• pp.343-347
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• 1988

The promoters of alkali-tolerant Bacillus sp. had been cloned in the promoter probe vector pPL703 and recombinant plasmid p-12 had been constructed. As a result of subcloning, two different promoters were found to exist in the cloned 2.9 kb promoter fragment and two recombinant plasmids p-l2B1 and p-l2B2, each harboring different promoter, were constructed. The promoter activity, which was expressed in the CAT specific activity, of p-l2B1 was 7 times higher than that of p-l2B2. The promoter activity as a function of growth revealed that both promoters of p-l2B1 and p-l2B2 were expressed after the late logarithmic growth phase and repressed in the presence of 1.0％ (w/v) glucose.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.70-79
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• 2014

In an attempt to develop a variety of expression vector systems for Corynebacterium glutamicum, six types of promoters, including $P_{tac}$, $P_{sod}$, $P_{sod}$ with a conserved Shine-Dalgarno (SD) sequence from C. glutamicum, $P_{ilvC}$, $P_{ilvC}$ with a conserved SD-1 ($P_{ilvC-M1}$), and $P_{ilvC}$ with a conserved SD-2 ($P_{ilvC-M2}$), were cloned into a modified shuttle vector, pCXM48. According to analysis of promoter strength by quantitative reverse transcription PCR, $P_{sod}$ and $P_{sod-M}$ were superior to tac and ilvC promoters in terms of transcription activity in C. glutamicum. All of the promoters have promoter activities in Escherichia coli, and $P_{sod-M}$ displayed the highest level of transcriptional activity. The protein expression in constructed vectors was evaluated by measuring the fluorescence of green fluorescent protein (GFP) and SDS-PAGE. C. glutamicum harboring plasmids showed GFP fluorescence with an order of activity of $P_{ilvC}$ > $P_{ilvC-M1}$ > $P_{sod}$ > $P_{ilvC-M2}$ > $P_{sod-M}$, whereas all plasmids except pCSP30 with $P_{sod}$ displayed fluorescence activities in E. coli. Of them, the strongest level of GFP was observed in E. coli with $P_{sod-M}$, and this seems to be due to the introduction of the conserved SD sequence in the translational initiation region. These results demonstrate that the expression vectors work well in both C. glutamicum and E. coli for the expression of target proteins. In addition, the vector systems harboring various promoters with different strengths, conserved SD sequences, and multiple cloning sites will provide a comfortable method for cloning and gene expression, and consequently contribute to the metabolic engineering of C. glutamicum.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1586-1592
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• 2017

Some promoters were isolated and characterized from the genome of Leuconostoc mesenteroides SY2, an isolate from kimchi, a Korean traditional fermented vegetable. Chromosomal DNA of L. mesenteroides SY2 was digested with Sau3AI and ligated with BamHI-cut pBV5030, a promoter screening vector containing a promoterless cat-86. Among E. coli transformants (TFs) resistant against Cm (chloramphenicol), 17 were able to grow in the presence of $1,000{\mu}g/ml$ Cm and their inserts were sequenced. Transcription start sites were examined for three putative promoters (P04C, P25C, and P33C) by primer extension. Four putative promoters were inserted upstream of a promoterless ${\alpha}$-amylase reporter gene in $pJY15{\alpha}$. ${\alpha}$-Amylase activities of E. coli TFs containing $pJY15{\alpha}$ (control, no promoter), $pJY03{\alpha}$ ($pJY15{\alpha}$ with P03C), $pJY04{\alpha}$ (with P04C), $pJY25{\alpha}$ (with P25C), and $pJY33{\alpha}$ (with P33C) were 66.9, 78.7, 122.1, 70.8, and 99.3 U, respectively. Cells harboring $pJY04{\alpha}$ showed 1.8 times higher activity than the control. Some promoters characterized in this study might be useful for construction of food-grade expression vectors for Leuconostoc sp. and related lactic acid bacteria.

• Journal of Convergence for Information Technology
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• v.11 no.2
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• pp.107-116
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• 2021

In this study, an analytical method was developed for estradiol-17��, testosterone, and progesterone, which are growth promoters as veterinary drug residues in animal food. The analytes were separated using liquid chromatography, and was injected into a mass spectrometer through the electrospray ionization(ESI) process and detected in multiple reaction monitoring(MRM) mode. As the method was validated by CODEX CAC/GL 71-2009 guideline, it met the acceptable. The analysis of beef, pork, and chicken distributed in Korea was conducted with an established method to confirm the applicability of the actual sample. It was confirmed that the developed method can be quickly and reliably in the analysis for the growth promoters identified in domestic distributed livestock products. Through subsequent research, a highly utilized analysis method will be completed if the number of growth promoters is expanded based on the method and the simultaneous analytical method is established by including all of it.

• Korean Journal of Air-Conditioning and Refrigeration Engineering
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• v.10 no.2
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• pp.193-201
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• 1998

A heat exchanging system employing the impinging air jet is still widely used In the various fields due to its inherent merits that include the easiness in engineering applications and the high heat and/or mass transfer characteristics. The purpose of this study is to investigate the enhancement of heat transfer and flow characteristics by placing a turbulence promoters in front of heat exchanging surface. In this study, a series of circular rods are placed at the upstream of a flat plate heat exchanger that is located at potential core region(H/W=2) of a two-dimensional impinging air jet. Heat transfer enhancement is achieved by inserting turbulence promoter that results in the flow acceleration and disturbance of boundary layer. The average Nusselt number of the flat plate with the turbulence promoters is found to be around 1.42 times higher than that of the flat plate without the turbulence promoters. Based on the results of flow visualization with a smoke wire, it is confirmed that the heat transfer enhancement is caused by the flow separation and disturbance of boundary layer by inserting the turbulence promoter.

• Environmental Mutagens and Carcinogens
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• v.25 no.2
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• pp.66-70
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• 2005

Telomerase, a specialized RNA-directed DNA polymerase that extends telomeres of eukaryotic chromosomes, has activity in most malignant tumors and provides a mechanism for the unlimited potential for division of neoplastic cells. Although telomerase is known to be a regulated enzyme, the factors and mechanisms involved in telomerase regulation are not well understood. In the present study, we compared the effect of 12-O­tetradecanoyl-phorbol-13-acetate (TPA) and non-phorbol ester tumor promoters such as okadaic acid, anthralin and benzoyl peroxide on the expression of telomerase in the mouse skin carcinogenesis system, a well characterized model for studying pre-malignant and malignant progression. We found that most early papillomas harvested after 10 weeks of TPA promotion showed telomerase activity. Other papillomas harvested after 10 weeks of okadaic acid, anthralin and benzoyl peroxide promotion and after single treatment of DMBA only also showed telomerase activity, respectively. On the other hand, normal and all skins surrounded by papillomas harvested after 10 weeks of these promoters has no telomerase activity. Taken together these results, there appears to be no clear association between the level of telomerase activity and protein phosphorylation in mouse skin papillomas and telomerase may be useful as bio-markers in early detection of tumors.