• Title/Summary/Keyword: high-level expression

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Construction of Yeast Vectors Potentially Useful for Expression of Eukaryotic Genes as ${\beta}$-galactosidase Fusion Proteins

  • Chung, Kyung-Sook;Choi, Won-Ja;Lee, Hee-Won;Kim, Kyu-Won;Yoo, Hyang-Sook
    • BMB Reports
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    • v.29 no.4
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    • pp.359-364
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    • 1996
  • By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6~7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

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Gene Expression Patterns of Spleen, Lung and Brain with Different Radiosensitivity in C57BL6 Mice

  • Majumder Md. Zahidur Rahman;Lee, Woo-Jung;Lee, Su-Jae;Bae, Sang-Woo;Lee, Yun-Sil
    • Journal of Radiation Protection and Research
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    • v.30 no.4
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    • pp.197-208
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    • 2005
  • Although little information is available on the underlying mechanisms, various genetic factors have been associated with tissue-specific responses to radiation. In the present study, we explored the possibility whether organ specific gene expression is associated with radiosensitivity using samples from brain, lung and spleen. We examined intrinsic expression pattern of 23 genes in the organs by semi-quantitative RT-PCR method using both male and female C57BL/6 mice. Expression of p53 and p21, well known factors for governing sensitivity to radiation or chemotherapeutic agents, was not different among the organ types. Both higher expression of sialyltransferase, delta7-sterol reductase, leptin receptor splice variant form 12.1, and Cu/Zn superoxide dismutase (SOD) and lower expression of alphaB crystalline were specific for spleen tissue. Expression level of glutathione peroxidase and APO-1 cell surface antigen gene in lung tissue was high, while that of Na, K-ATPase alpha-subunit, Cu/ZnSOD, and cyclin G was low. Brain, radioresistant organ, showed higher expressions of Na, K-ATPase-subunit, cyclin G, and nucleolar protein hNop56 and lower expression of delta7-sterol reductase. The result revealed a potential correlation between gene expression patterns and organ sensitivity, and Identified genes which might be responsible for organ sensitivity.

DEP Domain Containing 1 is a Novel Diagnostic Marker and Prognostic Predictor for Hepatocellular Carcinoma

  • Yuan, Sheng-Guang;Liao, Wei-Jia;Yang, Jian-Jun;Huang, Guo-Jin;Huang, Zhao-Quan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10917-10922
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    • 2015
  • Background: This study was conducted to determine DEPDC1 expression in hepatocelluar carcinomas (HCCs) and to reveal its potential role in diagnosis and prognosis of affected patients. Materials and Methods: DEPDC1 expression at the mRNA level was detected by quantitative real-time PCR (qRT-PCR) in 205 cases of HCC and paired adjacent normal liver tissues, and by semi-quantitative RT-PCR in 20 cases. Survival curves were obtained by using Kaplan-Meier method and Log-rank test. Independent predictors associated with regard to disease free survival (DFS) and overall survival (OS) were identified using the Cox proportional hazard model. Results: High DEPDC1 mRNA levels were detected in 144 out of 205 cases (70.24%) of HCC, significantly associated with clinicopathological parameters, including tumor size (${\geq}4cm$), alpha-fetoprotein (${\geq}100ng/ml$), B-C of BCLC stage and recurrence. Kaplan-Meier survival analysis revealed that HCC patients with high DEPDC1 expression had poor OS and DFS. Multivariate analysis demonstrated that high DEPDC1 expression was an independent predictor for OS (HR=1.651; 95% 95%CI, 1.041-2.617; p=0.033) and DFS (HR=1.583; 95%CI, 1.01-2.483; p=0.045). Conclusions: Our results indicate DEPDC1 might be a novel diagnostic marker and an independent prognostic predictor for HCC patients.

Toxicogenomics Study on Carbon Tetrachloride-induced Hepatotoxicity in Mice

  • Jeong, Sun-Young;Lim, Jung-Sun;Hwang, Ji-Yoon;Park, Han-Jin;Cho, Jae-Woo;Song, Chang-Woo;Kim, Yang-Seok;Lee, Wan-Seon;Moon, Jin-Hee;Han, Sang-Seop;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.275-280
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    • 2005
  • Carbon tetrachloride ($CCl_4$) is well known hepatotoxicant. Its overdose cause severe centrilobular hepatic necrosis in human and experimental animals. We administered $CCl_{4}$ at low (0.2 mL/kg p.o.) and high (2 mL/kg p.o.) doses to mice. Mice were sacrificed at 24 h after administration. We evaluated liver toxicity by serum AST and ALT level and by microscopic observation. Using cDNA chip, we conducted gene expression analysis in liver. Mean serum activities of the hepatocellular leakage enzymes, ALT and AST, were significantly increased compare to control, respectively, in the low and high dose groups. H&E evaluation of stained liver sections revealed $CCl_{4}-related$ histopathological findings in mice. Moderate centrilobular hepatocellular necrosis was present in all $CCl_{4}$ treated mice. We found that gene expression pattern was very similar between low and high dose group. However, some stress related genes were differently expressed. These results could be a molecular signature for the degree of liver injury. Our data suggest that the degree of severity could be figure out by gene expression profiling.

A Study On Female high-School Students' Reactive Characteristics to PSCD and Their Parent-Child Communication (여고생의 동그라미중심 부모-자녀 그림(PSCD) 반응특성과 부모-자녀 의사소통에 관한 연구)

  • Kim, Gab-Sook;Jeon, Young-Sook
    • Korean Journal of Human Ecology
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    • v.16 no.5
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    • pp.921-932
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    • 2007
  • This study is to examine the relationship between their parent-child communication and their reactive characteristics to the PSCD(Parent-Self-Centered-Drawing) test of the Female high-School Students. The study subjects are 209 Female high-School Students, and research tools are PSCD test and a measure of parent-child communication. The data were analyzed with a paired t-test, one-way ANOVA and Scheffe's post-hoc test. Results are as follows. First, they perceived positively communication toward mother more than communication toward father. Second, as for communication toward father according to the subjects' reaction to the father figure, female students show difference in ommission of facial parts, ommission of body parts, balance of body parts, facial expression and positive and negative symbols. Third, as for communication toward mother according to the subjects' reaction to the mother figure, female students show difference in ommission of facial parts, ommission of body parts, balance of body parts, facial expression and negative symbols. Forth, according to the female students' reaction to the self figure, they show different levels of communication toward father and self in regard of facial expression and distance between the figures. On the other hand, they show different levels of communication toward mother with respect to ommission of facial parts, ommission of body parts, balance of body parts, facial expression and distance between the figures. The authors conclude that the PSCD is a useful tool for measuring the level of parent-child communication.

Parallel Regulation of Prolactin and c-fos Gene Expression by 17$\beta$-estradiol and Stress in the Mouse Pituitary

  • Kim, Ji-Eune;Ko, Ji-Yun;Kim, Young-il;Yoon, Yong-Dal;Cho, Byung-Nam
    • Animal cells and systems
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    • v.4 no.1
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    • pp.71-76
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    • 2000
  • The aim of this study was to investigate expression patterns of the prolactin (PRL) and c-fos genes by 17$\beta$-estradiol (17$\beta$-E) and stress in the mouse pituitary. In the pituitary, the levels of PRL mRNA were found high with some fluctuation at 30, 50, and 90 min whereas the levels of PRL mRNA were low at 120 min when ovariectomized female mice were injected with 17$\beta$-E or vehicle. PRL mRNA levels began to increase again at 4 h and remained high up to 24 h only in the 17$\beta$-E- treated mice. The overall changes in c-fos mRNA by 17$\beta$-E were very similar to those in PRL mRNA in the pituitary. Subsequent study revealed that these high initial levels of PRL and c-fos mRNAs were caused by stress during Injection, not by 17$\beta$-E, since vehicle injection alone into the ovariectomized mice could increase the levels of PRL and c-fos mRNAs. The stress-induced elevations of PRL and c-fos mRNAs were inhibited by bromocriptin, a dopamine agonist, suggesting that the dopaminergic system is involved in the action route of injection stress. In addition, the induced levels of c-fos mRNA by 17$\beta$-E and stress in the pituitary were very low compared with those in the uterus. The time course changes in c-fos mRNA level were different between the pituitary and uterus. Taken together, these data indicate that PRL and c-tos gene expression in the pituitary are regulated by 17$\beta$-E and stress in a parallel manner, supporting the notion that c-Fos plays a role in regulation of PRL gene expression.

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A review of gene selection methods based on machine learning approaches (기계학습 접근법에 기반한 유전자 선택 방법들에 대한 리뷰)

  • Lee, Hajoung;Kim, Jaejik
    • The Korean Journal of Applied Statistics
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    • v.35 no.5
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    • pp.667-684
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    • 2022
  • Gene expression data present the level of mRNA abundance of each gene, and analyses of gene expressions have provided key ideas for understanding the mechanism of diseases and developing new drugs and therapies. Nowadays high-throughput technologies such as DNA microarray and RNA-sequencing enabled the simultaneous measurement of thousands of gene expressions, giving rise to a characteristic of gene expression data known as high dimensionality. Due to the high-dimensionality, learning models to analyze gene expression data are prone to overfitting problems, and to solve this issue, dimension reduction or feature selection techniques are commonly used as a preprocessing step. In particular, we can remove irrelevant and redundant genes and identify important genes using gene selection methods in the preprocessing step. Various gene selection methods have been developed in the context of machine learning so far. In this paper, we intensively review recent works on gene selection methods using machine learning approaches. In addition, the underlying difficulties with current gene selection methods as well as future research directions are discussed.

Molecular Characterization of Ischemia-Responsive Protein 94 (irp94) Response to Unfolded Protein Responses in the Neuron

  • Kim Seung-Whan;Kwon Ki-Sang;Shin Kee-Sun;Kim Seung-Ho;Kwon O-Yu
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.81-89
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    • 2006
  • The ischemia-responsive 94 gene (irp94) encoding a 94 kDa endoplasmic reticulum resident protein was investigated its molecular properties associated with unfoled protein responses. First, the expression of irp94 mRNA was tested after the reperfusion of the transient forebrain ischemia induction at the central nervous system in three Mongolian gerbils. Second, irp94 expression in PC12 cells, which are derived from transplantable rat pheochromocytoma cultured in the DMEM media, was tested at transcriptional and translational levels. The half life of irp94 mRNA was also determined In PC12 cells. Last, the changes of irp94 mRNA expression were investigated by the addition of various ER stress inducible chemicals (A23187, BFA, tunicamycin, DTT and $H_2O_2$) and proteasome inhibitors, and heat shock. High level expression of irp94 mRNA was detected after 3 hours reperfusion in the both sites of the cerebral cortex and hippocampus of the gerbil brain. The main regulation of irp94 mRNA expression in PC 12 cells was determined at the transcriptional level. The half life of irp94 mRNA in PC12 cells was approximately 5 hours after the initial translation. The remarkable expression of irp94 mRNA was detected by the treatment of tunicamycin, which blocks glycosylation of newly synthesized polypeptides, and $H_2O_2$, which induces apoptosis. When PC12 cells were treated with the cytosol proteasome inhibitors such as ALLN (N-acetyl-leucyl-norleucinal) and MG 132 (methylguanidine), irp94 mRNA expression was increased. These results indicate that expression of irp94 was induced by ER stress including oxidation condition and glycosylation blocking in proteins. Expression of irp94 was increased when the cells were chased after heat shock, suggesting that irp94 may be involved in recovery rather than protection against ER stresses. In addition, irp94 expression was remarkably increased when cytosol proteasomes were inhibited by ALLN and MG 132, suggesting that irp94 plays an important role for maintaining the ERAD (endoplasmic reticulum associated degradation) function.

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A study on the relationship between personality and the factors for clothing selection among the high school girls (여고생의 성격특색과 의복선택 요인과의 상관관계 연구 - 서울 시내 여고생을 중심으로 -)

  • 정하신
    • Journal of the Korean Home Economics Association
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    • v.23 no.4
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    • pp.1-7
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    • 1985
  • The aim of this study is to find out the relation-ship between personality and te factors for clothing selection. For the test, 396 high school girl students in Seoul were selected, respectively. The general personality test sheets by Kim Giseok and the questionnaire besed on Park Eunju's study on the factors for clothing selection were given to the sample group. RESULTS : 1. Behavior of clothing selection according to expression of individuality is sign ificantly correlated with ascendancy and sociability at the level of .001, and with emotional stability at .50. 2. Behavior of clothing selection according to utility is significantly correlated with resopnsibility and emotional stability at the level of .01. 3. Behavior of clothing selection according to economy is significantly correlated with ascendancy and self-confidence at the level of .01, and with responsibility and sociablity at .05. 4. Behavior of clothing selection according to modesty is significantly correlated with ascendancy and sociability at level of .001, and with self-confidence at .01. 5. Behavior of clothing selection according to conformity is significantly correlated with ascendancy, emothional stability, and self-confidence at the level of .001, and with sociability at .05.

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Assessment of Bile Salt Effects on S-Layer Production, slp Gene Expression and, Some Physicochemical Properties of Lactobacillus acidophilus ATCC 4356

  • Khaleghi, M.;Kermanshahi, R. Kasra;Yaghoobi, M.M.;Zarkesh-Esfahani, S.H.;Baghizadeh, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.749-756
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    • 2010
  • In many conditions, bacterial surface properties are changed as a result of variation in the growth medium and conditions. This study examined the influence of bile salt concentrations (0-0.1%) on colony morphotype, hydrophobicity, $H_2O_2$ concentration, S-layer protein production, and slpA gene expression in Lactobacillus acidophilus ATCC 4356. It was observed that two types of colonies (R and S) were in the control group and the stress condition. When the bile level increased in the medium, the amount of S type was more than the R type. A stepwise increment in the bile concentration resulted in a stepwise decline in the maximum growth rate. The results showed that hydrophobicity was increased in 0.01%-0.02% bile, but it was decreased in 0.1% bile. Treatment by bile (0.01%-0.1%) profoundly decreased $H_2O_2$ formation. S-Layer protein and slpA gene expression were also altered by the stress condition. S-Protein expression was increased in the stress condition. The slpA gene expression increased in 0.01%-0.05% bile and it decreased in 0.1% bile. However, we found that different bile salt concentrations influenced the morphology and some surface properties of L. acidophilus ATCC 4356. These changes were very different in the 0.1% bile. It appears that the bacteria respond abruptly to 0.1% bile.