• Title/Summary/Keyword: Over-expression

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Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

  • Ji-Su Jun;Min-Joo Kim;KwangWon Hong
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.128-137
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    • 2023
  • Transcriptome analysis revealed that the sinR gene encoding a transition-state regulator of Bacillus pumilus, genetically close to B. subtilis, was expressed at high levels during growth. The sinR gene is the second gene of the sinIR operon consisting of three promoters and two structural genes in B. subtilis. This study used the sinIR promoter of B. subtilis DB104 to construct a recombinant protein expression system. First, the expression ability depending on the number of sinIR promoter was investigated using enhanced green fluorescent protein (eGFP). The expression level of eGFP was slightly higher when using two promoters (Psin2) than using original promoters. The Psin2 promoter was further engineered by modifying the repressor binding site and -35 and -10 regions. Shine-Dalgarno (SD) sequence of the sinI gene was modified to the consensus sequence. Finally, combining the engineered Psin2 promoter with the modified SD sequence increased the expression level of eGFP by about 13.4-fold over the original promoter. Our results suggest that the optimized sinIR promoter could be used as a novel tool for recombinant protein expression in B. subtilis.

Musical Therapeutic Approch for Improving Self-expression and Self-esteem of Persons with Chronic Mental Disorder (음악치료가 만성정신장애인의 자기표현 및 자아존중감 향상에 미치는 효과)

  • Kim, Kae-Won;Cheong, Kwang-Jo;Choi, Ae-Na
    • Journal of Families and Better Life
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    • v.29 no.3
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    • pp.43-57
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    • 2011
  • Chronic mental-handicapped people are lacking in non-verbal expression such as eye contact, intonation, voice volume, facial expression, and gesture as well as the contents of speech, speak with a monotonous voice, fail to be vivid and clear in voice, and have absence of expression, thereby bringing about difficulty even for social adjustment and about low self-esteem. Accordingly, the purpose of this study was to examine effectiveness for enhancing self-expression and self-esteem by applying music therapy to the chronic mental-handicapped. The subjects were the chronic mental-handicapped who receive rehabilitation service at the community rehabilitation center, and who have over 10 years in the duration of disease. 1he music therapy activity was progressed with totally 14 sessions during 7 weeks with twice a week. This study confirmed t-test that is verification of difference in the mean, in order to examine difference between before and after music therapy in self-expression and self-esteem of the chronic mental-handicapped, and researched into qualitative case. The findings are as follows. First, as a result of score in self-expression scale, the significant improvement was shown after music therapy compared to before music therapy. The significant difference was indicated in verbal self-expression, phonetic self-expression, and non-verbal self-expression, which are its sub-spheres. Thus, the conclusion was obtained as saying that music therapy is effective for enhancing self-expression. Second, as a result of score in self-esteem scale, the significant difference was shown after music therapy compared to before music therapy. Thus, the conclusion was obtained as saying that music therapy is effective for enhancing self-esteem. Through the above results, the music therapy showed effectiveness of self-expression ability and self-esteem in the chronic mental-handicapped at the community rehabilitation center, thereby having been confirmed to be possibly utilized as rehabilitation program for the social skill ability and the social adjustment of the chronic mental-handicapped.

Tiul1 and TGIF are Involved in Downregulation of $TGF{\beta}1$-induced IgA Isotype Expression

  • Park, Kyoung-Hoon;Nam, Eun-Hee;Seo, Goo-Young;Seo, Su-Ryeon;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.248-254
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    • 2009
  • [ $TGF-{\beta}1$ ]is well known to induce Ig germ-line ${\alpha}$ ($GL{\alpha}$) transcription and subsequent IgA isotype class switching recombination (CSR). Homeodomain protein TG-interacting factor (TGIF) and E3-ubiquitin ligases TGIF interacting ubiquitin ligase 1 (Tiul1) are implicated in the negative regulation of $TGF-{\beta}$ signaling. In the present study, we investigated the roles of Tiul1 and TGIF in $TGF{\beta}1$-induced IgA CSR. We found that over-expression of Tiul1 decreased $TGF{\beta}1$-induced $GL{\alpha}$ promoter activity and strengthened the inhibitory effect of Smad7 on the promoter activity. Likewise, overexpression of TGIF also diminished $GL{\alpha}$ promoter activity and further strengthened the inhibitory effect of Tiul1, suggesting that Tiul1 and TGIF can down-regulate $TGF{\beta}1$-induced $GL{\alpha}$ expression. In parallel, overexpression of Tiul1 decreased the expression of endogenous IgA CSR-predicitive transcripts ($GLT_{\alpha},\;PST_{\alpha},\;and\;CT_{\alpha}$) and $TGF{\beta}1$-induced IgA secretion, but not $GLT_{\gamma3}$ and IgG3 secretion. Here, over-expressed TGIF further strengthened the inhibitory effect of Tiul1. These results suggest that Tiul1 and TGIF act as negatively regulators in $TGF{\beta}1$-induced IgA isotype expression.

Gene Expression of Hyperthyroid Rats treated by Scrophularia buergeriana Miquel (현삼(玄蔘)이 갑상선기능항진증(甲狀腺機能亢進症) 유발(誘發) 흰쥐의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Cho, Chung-Sik;Kim, Dae-Bok;Kim, Cheol-Jung
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.9-19
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    • 2007
  • Objective : The study was done to investigate Scrophularia buergeriana Miquel's effects in the genomic level, by measuring gene expression in hyperthyroid induced rats using cDNA micro array. Methods : Spargue-Dawley rats were separated into three groups(each with 10 rats). Except normal group, the other two groups were treated with Sodium levothyroxine $160{\mu}g$/kg/days for 5 days by oral administration. After 2 hours of the last intake of Sodium levothyroxine on two experimental group, one group was treated Scrophularia buergeriana Miquel extract 1.0g/kg/days for 3 days. The other groups(normal group, administration group) was treated normal saline 1.0g/kg/days for 3 days. Gene expression of hyperthyroid rats were measured after medication of solid extract of Scrophularia buergeriana Miquel with cDNA microarray. Results : In thyroid tissue, the numbers of the genes showed over twice increase and decrease in the control group compared to the normal group were 296, in the Scrophularia buergeriana Miquel administration group compared to the normal group were 859 and in the hypothalamus, the numbers of the genes showed over twice increase and decrease in the control group compared to the normal group were 416, in the Scrophularia buergeriana Miquel administration group compared to the normal group were 391. Conclusion : According to the above results, it is suggested that Scrophulari buergeriana Miquel suppress hyperthyroidism effectively and regulate the gene expression in the thyroid and brain.

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Missing values imputation for time course gene expression data using the pattern consistency index adaptive nearest neighbors (시간경로 유전자 발현자료에서 패턴일치지수와 적응 최근접 이웃을 활용한 결측값 대치법)

  • Shin, Heyseo;Kim, Dongjae
    • The Korean Journal of Applied Statistics
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    • v.33 no.3
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    • pp.269-280
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    • 2020
  • Time course gene expression data is a large amount of data observed over time in microarray experiments. This data can also simultaneously identify the level of gene expression. However, the experiment process is complex, resulting in frequent missing values due to various causes. In this paper, we propose a pattern consistency index adaptive nearest neighbors as a method of missing value imputation. This method combines the adaptive nearest neighbors (ANN) method that reflects local characteristics and the pattern consistency index that considers consistent degree for gene expression between observations over time points. We conducted a Monte Carlo simulation study to evaluate the usefulness of proposed the pattern consistency index adaptive nearest neighbors (PANN) method for two yeast time course data.

X-linked Gene Expression Profiles by RNAi-Mediated BRCA1 Knockdown in MCF7 Cells

  • Song, Min-Ae;Park, Jung-Hoon;Ahn, Hee-Jeong;Ko, Jung-Jae;Lee, Su-Man
    • Genomics & Informatics
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    • v.3 no.4
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    • pp.154-158
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    • 2005
  • Germ-line mutations of the BRCA1 gene confer an increased risk for breast and ovarian cancers. BRCA1 in female cells is directly related with the maintenance of the inactive X chromosome (Xi). The effect by the loss of the BRCA1 function on the X chromosome gene expression remains unclear in cancer cells. We attempted to investigate the expression pattern of the X-linked genes by performing BRCA1 knockdown via RNA interference in the MCF7 breast cancer cell line. The transcriptional and translational levels of BRCA1 were decreased over 95% in the MCF7 cells after BRCA1 knockdown. The expression patterns of one hundred ninety X-linked genes were profiled by the X chromosome-specific cDNA arrays. A total of seven percent of the X-linked genes (14/190) were aberrantly expressed by over 2-fold in the MCF7-BRCA1 knockdown cells, which contained two up-regulated genes (2/190, 1 %) and 12 down-regulated genes (12/190, 6.3%). It is interesting that 72% of the aberrantly expressed X-linked genes were located on the Xq (10/14,) region. Our data suggests that BRCA1 may not be important to maintain X chromosome inactivation in cancer because the BRCA1 knockdown did increase the expression of the only one percent of X-linked genes in the human breast cancer cells.

Regulation of nsdD Expression in Aspergillus nidulans

  • Han, Kap-Hoon;Han, Kyu-Yong;Kim, Min-Su;Lee, Dong-Beom;Kim, Jong-Hak;Chae, Suhn-Kee;Chae, Keon-Sang;Han, Dong-Min
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.259-261
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    • 2003
  • The nsdD gene has been predicted to encode a GATA type transcription factor with the type IVb zinc finger DNA binding domain functions in activating sexual development of A. nidulans. In several allelic mutants of nsdD producing truncated NsdD polypeptides lacking the C-terminal zinc finger, the transcription level of nsdD gene was greatly increased. Also in an over-expressed mutant, the transcription under its own promoter was reduced. These results suggest that the expression of nsdD is negatively autoregulated. When the nsdD gene was over-expressed, cleistothecia were formed in excess amounts even in the presence of 0.6 M KC1 that inhibited sexual development of the wild type. Northern blot analysis revealed that the expression of nsdD was repressed by 0.6 M KC1. These results strongly suggest that the inhibition of sexual development by salts was carried out via the nsdD involved regulatory network.

Cellular zinc deficiency inhibits the mineralized nodule formation and downregulates bone-specific gene expression in osteoblastic MC3T3-E1 cells

  • Cho, Young-Eun;Kwun, In-Sook
    • Journal of Nutrition and Health
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    • v.51 no.5
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    • pp.379-385
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    • 2018
  • Purpose: Zinc (Zn) is an essential trace element for bone mineralization and osteoblast function. We examined the effects of Zn deficiency on osteoblast differentiation and mineralization in MC3T3-E1 cells. Methods: Osteoblastic MC3T3-E1 cells were cultured at concentration of 1 to $15{\mu}M$ $ZnCl_2$ (Zn- or Zn+) for 5, 15 and 25 days up to the calcification period. Extracellular matrix mineralization was detected by staining Ca and P deposits using Alizarin Red and von Kossa stain respectively, and alkaline phosphatase (ALP) activity was detected by ALP staining and colorimetric method. Results: Extracellular matrix mineralization was decreased in Zn deficiency over 5, 15, and 25 days. Similarly, staining of ALP activity as the sign of an osteoblast differentiation, was also decreased by Zn deficiency over the same period. Interestingly, the gene expression of bone-related markers (ALP, PTHR; parathyroid hormone receptor, OPN; osteopontin, OC; osteocalcin and COLI; collagen type I), and bone-specific transcription factor Runx2 were downregulated by Zn deficiency for 5 or 15 days, however, this was restored at 25 days. Conclusion: Our data suggests that Zn deficiency inhibits osteoblast differentiation by retarding bone marker gene expression and also inhibits bone mineralization by decreasing Ca/P deposition as well as ALP activity.

Isolation and Characterization of Rice OsHRL Gene Related to Bacterial Blight Resistance

  • Park, Sang-Ryeol;Moon, Seok-Jun;Shin, Dong-Jin;Kim, Min-Gab;Hwang, Duk-Ju;Bae, Shin-Chul;Kim, Jeong-Gu;Yi, Bu-Young;Byun, Myung-Ok
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.417-420
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    • 2010
  • The expression of HR-like lesion inducing gene of Oryza sativa (OsHRL) was slightly increased by Xanthomonas oryzae pv. oryzae (Xoo) infection. Transgenic rice plants over-expressing OsHRL gene were challenged with Xoo and the development of disease symptoms were examined to investigate the effect of OsHRL gene expression on plant defense responses. The over-expression of OsHRL increased disease resistance against Xoo compared with wild type plants.

Overexpression of FGFR3 mRNA and Mutational Analysis of FGFR3 Gene in Hepatocellular Carcinoma (간암에서 FGFR3 mRNA의 과발현과 FGFR3 유전자의 돌연변이 분석)

  • Chang, Young Gyoon;Bae, Hyun Jin;Nam, Suk Woo
    • YAKHAK HOEJI
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    • v.56 no.6
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    • pp.352-357
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    • 2012
  • FGFR3 is a member of the fibroblast growth factor receptor family which interacts with fibroblast growth factors, setting in motion a cascade of downstream signals, ultimately influencing mitogenesis and differentiation. This particular family member binds acidic and basic fibroblast growth hormone and plays a role in bone development and maintenance. Accumulated evidence suggests that aberrant regulation of FGFR3 and genetic alterations are implicated in the development and progression of various cancers. Despite a high incidence of FGFR3 over-expression, no such investigation has been performed in hepatocellular carcinoma. Thus, we investigated genetic alterations of the FGFR3 gene in 73 cases of hepatocellular carcinoma by single-strand conformational polymorphism (SSCP) and sequencing. One silent mutation (A369A) was found in the extracellular domain of FGFR3, and one genetic alteration in the immunoglobulin-like III domain of FGFR3 appeared to be polymorphism. Taken together, we concluded that over-expression of FGFR3 in hepatocellular carcinoma is not associated with genetic alterations of FGFR3 gene, and we suggest that there could be another underlying mechanism of aberrant FGFR3 expression in hepatocellular carcinoma.