• Title/Summary/Keyword: Promoter analysis

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Molecular Mechanism of Photic-Entrainment of Chicken Pineal Circadian Clock

  • Okano, Toshiyuki;Fukada, Yoshitaka
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.25-28
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    • 2002
  • The chicken pineal gland has been used for studies on the circadian clock, because it retains an intracellular phototransduction pathway regulating the phase of the intrinsic clock oscillator. Previously, we identified chicken clock genes expressed in the gland (cPer2, cPer3, cBmal1, cBmal2, cCry1, cCry2, and cClock), and showed that a cBMALl/2-cCLOCK heteromer acts as a regulator transactivating cPer2 gene through the CACGTG E-box element found in its promoter. Notably, mRNA expression of cPer2 gene is up-regulated by light as well as is driven by the circadian clock, implying that light-dependent clock resetting may involve the up-regulation of cPer2 gene. To explore the mechanism of light-dependent gene expression unidentified in animals, we first focused on pinopsin gene whose mRNA level is also up-regulated by light. A pinopsin promoter was isolated and analyzed by transcriptional assays using cultured chicken pineal cells, resulting in identification of an 18-bp light-responsive element that includes a CACGTG E-box sequence. We also investigated a role of mitogen-activated protein kinase (MAPK) in the clock resetting, especially in the E-box-dependent transcriptional regulation, because MAPK is phospholylated (activated) in a circadian manner and is rapidly dephosphorylated by light in the gland. Both pulldown analysis and kinase assay revealed that MAPK directly associates with BMAL1 to phosphorylate it at several Ser/Thr residues. Transcriptional analyses implied that the MAPK-mediated phosphorylation may negatively regulate the BMAL-CLOCK-dependent transactivation through the E-box. These results suggest that the CACGTG E-box serves not only as a clock-controlled element but also as a light-responsive element.

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Association Analyses of ST8SIA2 Genetic Polymorphisms with Schizophrenia in the Korean Population (한국인 조현병 환자에서 ST8SIA2 유전자 다형성의 연합 연구)

  • Yoo, Jae-Hyun;Ryu, Seung-Hyong;Cho, Eun-Young;Huh, Ik-Soo;Park, Tae-Sung;Lee, Yu-Sang;Kwon, Jun-Soo;Hong, Kyung-Sue
    • Korean Journal of Biological Psychiatry
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    • v.19 no.3
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    • pp.140-145
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    • 2012
  • Objectives : ST8SIA2 (ST8 alpha-N-acetyl-neuraminide alpha-2, 8-sialyltransferase 2 gene) is located at 15q26, a susceptibility locus for schizophrenia. Some previous research had indicated that single-nucleotide polymorphisms (SNPs) in the promoter region of ST8SIA2 were associated with schizophrenia in Japanese and Chinese populations. We investigated the association between SNPs in the promoter region of ST8SIA2 and schizophrenia in the Korean population. Methods : The study subjects were 190 Korean patients with schizophrenia and 190 healthy controls. We performed allelic, genotypic, and haplotypic association analyses for rs3759916, rs3759915 and rs3759914 of ST8SIA2. All genotypes were determined by direct sequencing. Results : In the genotype-based analysis, rs3759914 showed a nominally significant association with schizophrenia under recessive genotypic model (p = 0.047). However, this association did not remain statistically significant after correction for multiple testing. Both allelic and haplotype analyses did not show any significant association. Conclusions : These findings suggest that ST8SIA2 does not play a major role in the susceptibility to schizophrenia in the Korean population. Further studies with a larger number of subjects are required to definitively rule out minor effects of this gene on schizophrenia vulnerability.

Comparative Analysis of Customer Feedback Metrics for Improving Predictability of Customer Loyalty (고객 충성도 예측력 제고를 위한 측정방법(CFM) 비교연구)

  • Kim, Joung-rae;Chung, Byoung-gyu
    • Journal of Venture Innovation
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    • v.1 no.2
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    • pp.61-73
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    • 2018
  • Many studies have focused on improving the predictability of influential variables in the area of customer services. For a long time, customer satisfaction index(CSI) were the best matrics of customer loyalty. In 2003, net promoter score(NPS) was introduced and in many cases this matrics was more predictive than CSI. In 2010, customer effort score(CES) was introduced and in the service areas this matrics was more predictive than CSI and NPS. This study compared with 3 matrics based on the criteria which composed of basic assumptions, time perspective, measuring items, objectives of matrics, application areas, and limitations of application. The dominant matric which was applied in all functions and industries was not exited. For the purpose of improving predictability, single or compound matrics are useful according to industries and usage.

Expression of the Apx Toxins of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae and Its Induction of Immune Response in Mice

  • Park Seung-Moon;Choi Eun-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yoo Han-Sang;Choi Woo Bong;Park Bong-Kyun;Kim Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.362-366
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    • 2005
  • Actinobacillus pleuropneumoniae is an important pig pathogen, which is responsible for swine pleuropneumonia, a highly contagious respiratory infection. To develop subunit vaccines for A. pleuropneumoniae infection, the Apx toxin genes, apxI and apxII, which are thought to be important for protective immunity, were expressed in Saccharomyces cerevisiae, and the induction of immune responses in mice was examined. The apxI and apxII genes were placed under the control of a yeast hybrid ADH2-GPD promoter (AG), consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. Western blot analysis confirmed that both toxins were successfully expressed in the yeast. The ApxIA and ApxIIA-specific IgG antibody response assays showed dose dependent increases in the antigen-specific IgG antibody titers. The challenge test revealed that ninety percent of the mice immunized with ApxIIA or a mixture of ApxIA and ApxIIA, and sixty percent of mice immunized with ApxIA survived, while none of those in the control groups survived longer than 36 h. These results suggest that vaccination of the yeast ex­pressing the ApxI and ApxII antigens is effective for the induction of protective immune responses against A. pleuropneumoniae infections in mice.

Presence of Transgenic Genes and Proteins in Commercial Soybean Foods from Mexican Grocery Stores

  • Cruz-Flores, Yendi Arely;Rodriguez-Herrera, Raul;Aguilar-Gonzalez, Cristobal Noe;Contreras-Esquivel, Juan Carlos;Reyes-Vega, Maria de la Luz
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1092-1096
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    • 2008
  • Commercial food products from major cities of Coahuila, Mexico were screened to identify residues of transgenic deoxyribonucleic acid (DNA) and/or proteins. After performed, an inventory on all products that contained a soybean-based ingredient in a commercial grocery store in the city of Saltillo, Coahuila, Mexico, 245 food products were identified and grouped in 15 classes according to the soybean ingredient as well as the manufacturing process used for their elaboration. Similar sampling was made for the different food classes in the cities of Monclova, Piedras Negras, and Torreon. A total of 88 samples were analyzed and DNA was extracted by the hexadecyltrimethyl-ammonium bromide (CTAB) technique with slight modification to obtain better DNA quality (1). In addition, segments of the transgenic genes one that codifies for 5-enolpyruvylshikimate-3-phosphate synthase (epsps), cry 1A, and the cauliflower mosaic virus (CaMV) promoter were amplified using polymerase chain reaction (PCR). The transgenic proteins 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) and insecticidal crystal protein (Cry 1Ab/Ac) were identified using double antibody sandwich-enzymatic linked immunoassay analysis (DAS-ELISA). Presence of transgenic genes and/or proteins was identified in 35.3% of the commercial products samples.

Valproic Acid Induces Transcriptional Activation of Human GD3 Synthase (hST8Sia I) in SK-N-BE(2)-C Human Neuroblastoma Cells

  • Kwon, Haw-Young;Dae, Hyun-Mi;Song, Na-Ri;Kim, Kyoung-Sook;Kim, Cheorl-Ho;Lee, Young-Choon
    • Molecules and Cells
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    • v.27 no.1
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    • pp.113-118
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    • 2009
  • In this study, we have shown the transcriptional regulation of the human GD3 synthase (hST8Sia I) induced by valproic acid (VPA) in human neuroblastoma SK-N-BE(2)-C cells. To elucidate the mechanism underlying the regulation of hST8Sia I gene expression in VPA-stimulated SK-N-BE(2)-C cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5'-flanking region of the hST8Sia I gene by the transient expression method showed that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-${\kappa}B$, functions as the VPA-inducible promoter of hST8Sia I in SK-N-BE(2)-C cells. Site-directed mutagenesis and electrophoretic mobility shift assay indicated that the NF-${\kappa}B$ binding site at -731 to -722 was crucial for the VPA-induced expression of hST8Sia I in SK-N-BE(2)-C cells. In addition, the transcriptional activity of hST8Sia I induced by VPA in SK-N-BE(2)-C cells was strongly inhibited by SP600125, which is a c-Jun N-terminal kinase (JNK) inhibitor, and $G{\ddot{O}}6976$, which is a protein kinase C (PKC) inhibitor, as determined by RT-PCR (reverse transcription-polymerase chain reaction) and luciferase assays. These results suggest that VPA markedly modulated transcriptional regulation of hST8Sia I gene expression through PKC/JNK signal pathways in SK-N-BE(2)-C cells.

Development of Transgenic Soybean Using Agrobacterium tumefaciens (Agrobacterium tumefaciens을 이용한 대두 형질전환체 개발)

  • Cho, Mi-Ae;Choi, Dong-Woog;Liu, Jang-Ryol;Clemente Tom;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.255-259
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    • 2004
  • Agrobacterium tumefaciens-mediated cotyledonary node transformation was used to produce transgenic soybean. Cotyledonary node explants of three cultivars and one genotype were co-cultivated with strains Agrobacterium (LBA4404, GV3101, EHA101, C58) containing the binary vectors (pCAMBIA3301 and pPTN289) carrying with CaMV 35S promoter-GUS gene as reporter gene and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selectable marker. There was a significant difference in the transformation frequency depend on bacteria strain. The EHA101 strain of the bacterial strains employed gave the maximum efficiency (3.6%). One hundred-six lines transformed showed the resistance in glufosinate. Histochemical GUS assay showed that at least 11 plants transformed with the GUS gene were positive response. The soybean transformants were obtained from the Thorne (5 plants), 1049 (5 plants) and Bakun (1 plant), respectively. Southern blot analysis and leaf painting assay revealed that the GUS and bar gene segregated and expressed in their progeny.

Construction and Characterization of Transformed Insect Cells Expressing Baculovirus Very Late Factor in an Infection-Independent Manner

  • Park, Hye-Jin;Lee, Kwang-Sik;Cho, Eun-Sook;Yun, Eun-Young;Kang, Seok-Woo;Kim, Keun-Young;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.19-26
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    • 2001
  • Transformed Spodoptera frugiperda (Sf9) cells expressing baculovirus very late factor (VLF-1) were constructed by using Autograha nuclear polyhedrosis virus (AcNPV) immediate earthy gene (ie1). Neomycin-resistance gene as a selectable marker was introduced under the control of AcNPV ie1 promoter, and Bombyx mori nuclear polyhedrosis (BmNPV-K1) vlf-1 gene was introduced under the control of the Drosophila heat shock protein gene (hspr70) promoter to yield dual expression plasmid with two independent transcription units. It was transfected into Sf9 cells and cell clones expressing vlf-1 were selected by G4l8 treatment. Genomic DNA from transformed cells was isolated and integration of AcNPV iel harboring vlf-1 was confirmed by PCR using AcNPV iel-specific primers and Southern blot analysis. The transformed cells expressing VLF-1 in an infection-independent manner expressed foreign gene product of recombinant baculovirus in the earlier stage of infection compared with control Sf9 cells. These results suggest the possible to develop highly efficient transformed insect cells for baculovirus expression vector system.

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Attitude toward Health behaviors in Student Nurses (간호대학생의 건강행위에 대한 태도 조사연구)

  • Jung, Moon-Hee;Cho, Yoo-Hyang;Kim, Myung-Soon;Kim, Hyun-Li
    • Research in Community and Public Health Nursing
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    • v.13 no.4
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    • pp.826-835
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    • 2002
  • Health perception is a very important issue for student nurses to be future health professionals. The concept of health has changed with the emphasis of 'health promotion', and thus nurses now have a demanded role as a health promoter. According to health promotion theories, health-promoting behaviors are affected by cognitive variables, and human behaviors are related to thoughts and perception. This study was conducted to recognize the relationships of attitude toward health behaviors to other health related variables. The data was collected using a self administered survey. The instrument used in this study was attitude of health behavior scales that were developed by Okayama medical school. The subjects were 512 student nurses recruited from 1 college and 3 universities. The data was analyzed using SPSS pc program with mean, correlation. and multiple regression technique. The results were as follows: 1. The mean score of attitude of health behaviors of the student nurses was 8.11, showing a relatively high level. A high score of health belief was reported in 'self-responsible type' with the score of 4.80, and a high score of 'think for health or disease' was reported in passive type with the score of 3.12. 2. Specifically, there were significant positive correlations between the level of 'attitude of health behaviors' and other health related variables such as health perception, think for health or disease, and age. 3. Stepwise multiple regression analysis revealed that active thought, leisure, self-responsible perception and depression accounted for 13%of variance of attitude toward health behaviors. In conclusion, although student nurses certainly perceived the performance of health behavior, they need to learn in more systematical way in order to be a better health promoter, one of the role of nurses. The results of the study suggest that further studies need to be done on attitude toward health behaviors and performance of health behavior by student nurses.

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Comparative Evaluation on Qualitative PCR using Different Extraction Methods for Nucleic Acids on Soybean and Corn Processed Foods (대두 및 옥수수 가공식품에서 유전자재조합체(GMO)의 정성 PCR분석을 위한 핵산 추출방법별 비교)

  • 김영찬;이철수;황순욱;김성조;이영옥;윤성원;서정화;남용석
    • Journal of Food Hygiene and Safety
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    • v.18 no.1
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    • pp.6-13
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    • 2003
  • Various kinds of genetically modified organisms (GMO) and processed foods have been developed during recent years. Genetically modified organisms can be classified into several groups as their development methods. Generally, GMO has three foreign DNA regions such as gene expression adjustment region(Promoter), termination region (terminator) and structure gene. Detection of these regions can be done particularly by polymerase chain reaction (PCR). PCR-based detection can virtually be performed for any GMO within short of time. The most important prerequisite for the application of PCR-based detection is to decide abstraction method of efficient nucleic acids. Specially, in the case of processed food, because nucleic acids of foodstuffs are damaged by heat treatment (sterilization), pressure and fermentation, DNA must be extracted ken the samples prior to PCR analysis. Although many DNA extraction protocols are available, they have rarely been compared in a comprehensive method. In this study low widely used commercial and non-commercial DNA extraction methods-DNeasy$^{TM}$, Wizard$^{TM}$, CTAB, phenol/chloroform system-were compared with respect to the quality and yield of nucleic acids and insertion genes.nes.