• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.190-194
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• 2004

The gene product of dctD (DctD) activates transcription from the dctA promoter regulatory region by the $\sigma^{54}$ -holoenzyme form ofRNA polymerase ($E\sigma^{54}$ ) in Rhizobium meliloti and R. leguminosarum. The Escherichia coli integration host factor (IHF) stimulated DctD-mediated activation from the dctA promoter regulatory region of R. leguminosarum but not R. meliloti. In the absence of UAS, IHF inhibited DctD-mediated activation from both of these promoter regulatory regions. IHF also inhibited activation from R. leguminosarum dctA by nitrogen regulatory protein C (NtrC), another activator of $E\sigma^{54}$ but not by one which lacks a specific binding site in this promoter regulatory region. IHF, however, stimulated NtrC-mediated activation from the R. meliloti dctA promoter. Upon removal of the UAS, IHF inhibited NtrC-mediated transcription activation from the R. meliloti dctA promoter regulatory region. These data suggest that IHF likely faciliates productive contacts between the activators NtrC or DctD and $E\sigma^{54}$ to stimulate activation from dctA promoter.

• Development and Reproduction
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• v.23 no.1
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• pp.55-61
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• 2019

To develop a promoter capable of driving transgene expression in non-model fish, we identified and characterized the muscle-specific alpha-actin gene in olive flounder, Paralichthys olivaceus (PoACTC1). The regulatory region of PoACTC1 includes putative regulatory elements such as a TATA box, two MyoD binding sites, three CArG boxes, and a CCAAT box. Microinjection experiments demonstrated that the regulatory region of PoACTC1, covering from -2,126 bp to +751 bp, just prior to the start codon, drove the expression of red fluorescent protein in developing zebrafish embryos and hatching olive flounder. These results suggest that the regulatory region of PoACTC1 may be useful in developing a promoter for biotechnological applications such as transgene expression in olive flounder.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1628-1633
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• 2001

A goat ${\beta}$-casein gene was cloned and sequenced. Our previous study had determined the nucleotide sequences of the 5' flanking region and the structural gene including all 9 exons. In the present study, investigations were done on the regulatory sequences in the 5' flanking region of the goat ${\beta}$-casein gene by aligning and comparing it with the same gene from other mammals. The results showed that -200/-1 bp of the 5' flanking sequences contained six conserved clusters, in which the sites of gene expression regulated by the transcription factor and hormone might exist. It showed that fourteen glucocorticoid receptor elements, two cAMP responsive elements, two SV40 virus enhancer core sequences, two OCT-1 binding elements and one CTF/NF-1 binding element were dispersed in the 5' flanking region of goat ${\beta}$-casein gene. Our findings are perhaps valuable for the elucidation of the molecular mechanisms that control the expression of the goat ${\beta}$-casein gene.

• Archives of Pharmacal Research
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• v.21 no.1
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• pp.35-40
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• 1998

Human cytochrome P450 4F2 shows high regioselectivity in hydroxylation of stearic acid and leukotriene $ B_4.$ As a first step of its regulation study, human cytochrome P450 4F2 genomic DNA was isolated from liver of a person who was administered clofibrate for 10 years. From Southern hybridization, restriction enzyme digestion and sequencing experiments, isolated genomic DNA fragment was found to contain around 32 Kb DNA and more than 20 Kb of $5^I$ end regulatory region. Sequences of the structural gene region revealed exon 1 and exon 2. Further regulation studies would elucidate the feedback mechanisms of the oxidative degradation of fatty acids, inflammatory response and the clearance of leukotriene B4 in the liver. Furthermore, regulation study of this gene could explain the species difference in responses to peroxisome proliferator and help in the safety evaluation of peroxisome proliferating chemicals to human being.

• Journal of Life Science
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• v.5 no.4
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• pp.162-169
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• 1995

The glpD genes encoding gly-3-p dehydrogenase is essential for the aerobic growth of E. coli on glycerol or gly-3-p. The glpE gene, the function of which is unknownm is transcribed divergently with respect to glpD gene. Expression of the adjacent but divergently transcribed glpD the glpE genes is positively regulated by the cAMP-CRP complex. In this study, for a precise investigation of the functional elements in the regulatory region for transcription activation by cAMP-CRP, deletion mutation have been introducted into the regulatory region. The effect of the deletion mutant on transcriptional regulation was tested in vivo by $\beta$-galctosidase activity. Deletion mutants in the regulatory region of glpD demonstrated that the presence of the CRP-binding site resulted in an sixfold increase in promoter activity. And also deletion mutants of glpE gene demonstrated that the presence of the CRP-binding site resulted in an eightfold increase in promoter activity. Insertion of 22 bp oligomer in the deletion mutants has shown that the CRP binding site is need for maximal expression of glpD and glpE genes. glpD and glpE gene, cAMP-CRP complex, deletion mutant, transcriptional regulation.

• Communications for Statistical Applications and Methods
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• v.25 no.4
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• pp.341-354
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• 2018

The one of the principles described in ICH E9 is that only results obtained from pre-specified statistical methods in a protocol are regarded as confirmatory evidence. However, in multi-regional clinical trials, even when results obtained from pre-specified statistical methods in protocol are significant, it does not guarantee that the test treatment is approved by regional regulatory agencies. In other words, there is no so-called global approval, and each regional regulatory agency makes its own decision in the face of the same set of data from a multi-regional clinical trial. Under this situation, there are two natural methods a regional regulatory agency can use to estimate the treatment effect in a particular region. The first method is to use the overall treatment estimate, which is to extrapolate the overall result to the region of interest. The second method is to use regional treatment estimate. If the treatment effect is completely identical across all regions, it is obvious that the overall treatment estimator is more efficient than the regional treatment estimator. However, it is not possible to confirm statistically that the treatment effect is completely identical in all regions. Furthermore, some magnitude of regional differences within the range of clinical relevance may naturally exist for various reasons due to, for instance, intrinsic and extrinsic factors. Nevertheless, if the magnitude of regional differences is relatively small, a conventional method to estimate the treatment effect in the region of interest is to extrapolate the overall result to that region. The purpose of this paper is to investigate the effects produced by this type of extrapolation via estimations, followed by hypothesis testing of the treatment effect in the region of interest. This paper is written from the viewpoint of regional regulatory agencies.

• Molecules and Cells
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• v.41 no.4
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• pp.342-350
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• 2018

Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the ${\beta}$-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a-GUS construct to rice cultivar 'Taichung 65' which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.

• Fisheries and Aquatic Sciences
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• v.12 no.2
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• pp.90-97
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• 2009

The cytoskeletal $\beta$-actin gene and its 5'-upstream region were isolated and characterized in the rockbream (Oplegnathus fasciatus). Complementary DNA of the rockbream $\beta$-actin represented a 1,125 bp of an open reading frame encoding 375 amino acids, and the rockbream $\beta$-actin cDNA and deduced amino acid sequences were highly homologous to those of other vertebrate orthologs. At the genomic level, the $\beta$-actin gene also exhibited an organization typical of vertebrate cytoskeletal actin genes (2,159 bp composed of five translated exons interrupted by four introns) with a conserved GT/AG exon-intron splicing rule. The putative non-translated exon predicted in the rockbream $\beta$-actin gene was much more homologous with those of teleostean $\beta$-actin genes than those of mammals. The 5'-upstream regulatory region isolated by genome walking displayed conserved and essential elements such as TATA, CArG and CAAT boxes in its proximal part, while several other immune- or stress-related motifs such as those for NF-kappa B, USF, HNF, AP-1 and C/EBP were in the distal part. Semi-quantitative RT-PCR assay results demonstrated that the rockbream $\beta$-actin transcripts were ubiquitously but different-tially expressed across the tissues of juveniles.

• Bulletin of the Korean Chemical Society
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• v.22 no.8
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• pp.903-908
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• 2001

The gene aeg-46.5, which is expressed under anaerobic condition, has putative triple -10 regions and four transcription start sites. The mRNA transcription level and its start point change depending on the aerobic/anaerobic growth condition. RNA polymerase and its regulatory proteins must choose which of three -10 region to use. The putative triple 10 region was mutated to make only one of them function with consensus -10 region sequence (TATAAT) and the other two as non-functional region. The results show that the second and third -10 regions are used for the aerobic/anaerobic expression. The third -10 region is responsible for the high aerobic to anaerobic switch ratio. This suggests that only the last two of the putative triple -10 region have functions on aeg-46.5 gene expression switch control. The phenotype of the mutated promoter was tested in the wild type cell and narL - cell. The results indicate that the control by NarL is independent from the selection of -10 region. The expression patterns on multi-copy plasmids and on single-copy chromosome were compared. These results show that the aerobic/anaerobic switch control of aeg-46.5 is through the choice of -10 region. The mechanism of choosing different -10 region remains to be seen.

• YAKHAK HOEJI
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• v.37 no.5
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• pp.504-510
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• 1993

Human insulin gene is consisted of the polymorphic region with the repeating units, the regulatory sequence, the structural gene including the intervening sequence, and 3'-flanking region. The polymerase chain reaction, which amplifies the target DNA between two specific primers, has been performed for the amplification of human insulin gene and simple one-step cloning of it into Escherichia coli. Out of 1727 nuceotides compared, only 4 sites were variable: 5'-regulatory region(G2101$\rightarrow$AGG); IVS I(T2401$\rightarrow$A); Exon II(C2411 deletion); IVS II(A2740 dejection). The variations at the G2101 and T2401 were the same as those found in one American allele. The other two variations were observed only in the specific Korean allele. And, the enzyme digestion patterns among normal, insulin dependent diabetes mellitus, and non-insulin dependent diabetes mellitus were the same. On the other hand, PCR method showed the possibility of the quickaccess for the polymorphic region in terms of the restriction fragment length of polymorphism.