• Title/Summary/Keyword: Expression rate

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Investigation of the Growth Rate Change in Recombinant BCG which was cloned Mycobacterium tuberculosis Adenylate Kinase Mutation Gene or Human Muscle-type Adenylate Kinase Synthetic Gene (결핵균 Adenylate Kinase 돌연변이 유전자와 Human Muscle-type Adenylate Kinase 합성 유전자를 형질전환한 BCG의 성장속도 변화 유무 조사)

  • Lee, Seung-Heon;Kim, Hyo-Joon;Park, Young-Kil;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.2
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    • pp.187-193
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    • 2006
  • Background : Normal cell proliferation and viability is strongly depends on the availability of metabolic energy and the maintenance of the appropriate adenylate-nucleotide pools. Hypothetically, changes in adenylate kinase (AK) expression could therefore be associated with adaptation to altered growth characteristics or inversely altered growth characteristics of proliferating cells could drive the changes in the metabolic profile. This study investigated whether the expression of either AK1 or a Mycobacterium tuberculosis adenylate kinase mutant which has the same catalytic activity of AK1 could affect the growth rate of slow-growing BCG. Method : Recombinant BCGs, which were cloned the human muscle-type adenylate kinase synthetic gene (AK1) and adenylate kinase mutation gene (AKmtDM) of Mycobacterium tuberculosis into the Mycobacterium/E.coli expression vectors, were constructed. Recombinant BCGs and wild-type BCG were cultured in 7H9 media and the optical density at 600nm was measured at intervals of 2-3 days. Result : There wasn't the growth rate change induced by AK1 or AKmtDM expression in recombinant BCGs. Conclusion : The expression of AK1 or Mycobacterium tuberculosis adenylate kinase mutant in BCG does not affect the growth rate of BCG.

Prognostic Significance of Expression of CD133 and Ki-67 in Gastric Cancer

  • Saricanbaz, Irem;Karahacioglu, Eray;Ekinci, Ozgur;Bora, Huseyin;Kilic, Diclehan;Akmansu, Muge
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.19
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    • pp.8215-8219
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    • 2014
  • CD133 is one of the most important stem cell markers in solid cancers and Ki-67 is a marker that reflects cell proliferation. The relationships between the expression of CD133 and Ki-67 and prognosis in gastric carcinoma are unknown and need exploring. We examined 50 gastric cancer patients retrospectively in the Radiation Oncology Department of the Faculty of Medicine, Gazi University. CD133 and Ki-67 expression was examined using immunohistochemical staining. The survival rate in patients with CD133 positive expression was significantly worse than that in the patients with negative expression (p=0.04). Expression of CD133 had a positive correlation with that of Ki-67 (r=0.350; p=0.014). Multivariate analysis revealed that the expression of CD133 was an independent prognostic factor in gastric cancer (p=0.02). Conclusion, expression of CD133 may be a useful prognostic marker in gastric cancer.

Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.110-117
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    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

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Root metabolic cost analysis for root plasticity expression under mild drought stress

  • Kano-Nakata, Mana;Mitsuya, Shiro;Inukai, Yoshiaki;Yamauchi, Akira
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.328-328
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    • 2017
  • Drought is a major limiting factor that reduces rice production and occurs often especially under recent climate change. Plants have the ability to alter their developmental morphology in response to changing environment, which is known as phenotypic plasticity. In our previous studies, we found that one chromosome segment substitution line (CSSL50 derived from Nipponbare and Kasalath crosses) showed no differences in shoot and root growth as compared with the recurrent genotype, Nipponbare under non-stress condition but showed greater growth responses compared with Nipponbare under mild drought stress condition. We hypothesized that reducing root respiration as metabolic cost, which may be largely a consequence of aerenchyma formation would be one of the key mechanisms for root plasticity expression. This study aimed to evaluate the root respiration and aerenchyma formation under various soil moisture conditions among genotypes with different root plasticity. CSSL50 together with Nipponbare and Kasalath were grown under waterlogged conditions (Control) and mild drought stress conditions (20% of soil moisture content) in a plastic pot ($11cm{\times}14cm$, ${\varphi}{\times}H$) and PVC tube ($3cm{\times}30cm$, ${\varphi}{\times}H$). Root respiration rate was measured with infrared gas analyzer (IRGA, GMP343, Vaisala, Finland) with a closed static chamber system. There was no significant difference between genotypes in control for shoot and root growth as well as root respiration rate. In contrast, all the genotypes increased their root respiration rates in response to mild drought stress. However, CSSL50 showed lower root respiration rate than Nipponbare, which was associated by higher root aerenchyma formation that was estimated based on internal gas space (porosity) under mild drought stress conditions. Furthermore, there were significant negative correlations between root length and root respiration rate. These results imply that reducing the metabolic cost (= root respiration rate) is a key mechanism for root plasticity expression, which CSSL50 showed under mild drought.

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The Expression of Hsp70 and GST Genes in Mytilus coruscus during Air Exposure and Starvation (공기노출 및 절식시기 동안 참담치, Mytilus coruscus 에서 Hsp70 및 GST 유전자 발현에 대한 연구)

  • Kim, Chul Won;Kang, Han Seung
    • The Korean Journal of Malacology
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    • v.32 no.2
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    • pp.73-81
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    • 2016
  • Heat shock proteins (HSPs), one of the most highly conserved groups of proteins characterized to date, play crucial roles in protecting cells against environmental stresses, such as heat shock, salinity and oxidative stress. The glutathione S-transferases (GST) have important role in detoxification of oxidative stress, environmental chemicals and environmental stress. GST mRNA expression have been used as biomarkers on environmental stress. The purpose of this study was to investigate the death rate and the gene expression of Hsp70 and GST during air exposure and starvation. Results showed that, the expression of Hsp70 mRNA was significantly changed in the experiment groups, such as air exposure and starvation. GST mRNA expression was significantly increased in the experimental group of starvation. These results suggest that Hsp70 and GST were played roles in biomarker gene on the air exposure and starvation.

북한산 국립공원의 식생군집형에 대하여

  • 송호경;이근복
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.23-33
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    • 1985
  • Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

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Comparative Whole Cell Proteomics of Listeria monocytogenes at Different Growth Temperatures

  • Won, Soyoon;Lee, Jeongmin;Kim, Jieun;Choi, Hyungseok;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.259-270
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    • 2020
  • Listeria monocytogenes is a gram-positive, facultative anaerobe food pathogen responsible for the listeriosis that mostly occurs during the low-temperature storage of a cold cut or dairy products. To understand the systemic response to a wide range of growth temperatures, L. monocytogenes were cultivated at a different temperature from 10℃ to 42℃, then whole cell proteomic analysis has been performed both exponential and stationary cells. The specific growth rate increased proportionally with the increase in growth temperature. The maximum growth rate was observed at 37℃ and was maintained at 42℃. Global protein expression profiles mainly depended on the growth temperatures showing similar clusters between exponential and stationary phases. Expressed proteins were categorized by their belonging metabolic systems and then, evaluated the change of expression level in regard to the growth temperature and stages. DnaK, GroEL, GroES, GrpE, and CspB, which were the heat&cold shock response proteins, increased their expression with increasing the growth temperatures. In particular, GroES and CspB were expressed more than 100-fold than at low temperatures during the exponential phase. Meanwhile, CspL, another cold shock protein, overexpressed at a low temperature then exponentially decreased its expression to 65-folds. Chemotaxis protein CheV and flagella proteins were highly expressed at low temperatures and stationary phases. Housekeeping proteins maintained their expression levels constant regardless of growth temperature or growth phases. Most of the growth related proteins, which include central carbon catabolic enzymes, were highly expressed at 30℃ then decreased sharply at high growth temperatures.

A Functional SNP in the MDM2 Promoter Mediates E2F1 Affinity to Modulate Cyclin D1 Expression in Tumor Cell Proliferation

  • Yang, Zhen-Hai;Zhou, Chun-Lin;Zhu, Hong;Li, Jiu-Hong;He, Chun-Di
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3817-3823
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    • 2014
  • Background: The MDM2 oncogene, a negative regulator of p53, has a functional polymorphism in the promoter region (SNP309) that is associated with multiple kinds of cancers including non-melanoma skin cancer. SNP309 has been shown to associate with accelerated tumor formation by increasing the affinity of the transcriptional activator Sp1. It remains unknown whether there are other factors involved in the regulation of MDM2 transcription through a trans-regulatory mechanism. Methods: In this study, SNP309 was verified to be associated with overexpression of MDM2 in tumor cells. Bioinformatics predicts that the T to G substitution at SNP309 generates a stronger E2F1 binding site, which was confirmed by ChIP and luciferase assays. Results: E2F1 knockdown downregulates the expression of MDM2, which confirms that E2F1 is a functional upstream regulator. Furthermore, tumor cells with the GG genotype exhibited a higher proliferation rate than TT, correlating with cyclin D1 expression. E2F1 depletion significantly inhibits the proliferation capacity and downregulates cyclin D1 expression, especially in GG genotype skin fibroblasts. Notably, E2F1 siRNA effects could be rescued by cyclin D1 overexpression. Conclusion: Taken together, a novel modulator E2F1 was identified as regulating MDM2 expression dependent on SNP309 and further mediates cyclin D1 expression and tumor cell proliferation. E2F1 might act as an important factor for SNP309 serving as a rate-limiting event in carcinogenesis.

Expression and Prognostic Implications of FOXO3a and Ki67 in Lung Adenocarcinomas

  • Liu, Hong-Bin;Gao, Xiang-Xiang;Zhang, Qing;Liu, Jian;Cui, Yuan;Zhu, Yan;Liu, Yi-Fei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1443-1448
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    • 2015
  • To investigate the significance of FOXO3a and Ki67 in human lung adenocarcinomas. Envision immunohistochemical staining and Western blotting were used to examine the protein expression of FOXO3a in 127 cases of human lung adenocarcinoma specimens. The positive rate in lung adenocarcinoma (55.9%) was lower than that in normal tissues (80%). We found that the expression of FOXO3a was closely related with the degree of differentiation, TNM staging, lymph node metastasis and survival. In addition, significant differences in the different pathological types of lung adenocarcinoma cases (P<0.01). The FOXO3a positive rate of the acini as the main type (APA) (86.7%) and the lepidic as the main type (LPA) (82.4%) was higher than the solid as the main type (SPA) (50.0%), the papilla as the main type (PPA) (42.9%) and the micropapilla as the main type (MPA) (9.4%). Moreover, the expression of FOXO3a was negatively related with Ki67 expression. Our results suggested that the expression of FOXO3a is closely correlated with the aggressiveness of lung adenocarcinoma. It was indicated that disregulation of FOXO3a might play key roles in the occurrence and development of lung a denocarcinoma and joint detection of the two markers might play an important role in diagnosing tumors.

Generalized BER Analysis of Arbitrary Rectangular QAM (임의의 사각형 QAM의 일반화된 비트 오율 분석)

  • Yoon Dong-Weon;Cho, Kyong-Kuk;Suh, Ki-Bum
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.27 no.10A
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    • pp.962-968
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    • 2002
  • Reliable high-speed data communications over insufficient channel bandwidth is one of the major challenges of harsh wireless environments that push the achievable spectral efficiency far below its theoretical limits. A Quadrature amplitude modulation (QAM) scheme is a userful modulation technique for achieving high data rate transmission without increasing the bandwidth of wireless communication systems. The exact general bit error rate (BER) expression of arbitrary rectangular quadrature amplitude modulation has not yet been derived. In this paper, a generalized closed-form expression for the BER performance of rectangular QAM with Gray code bit mapping is derived and analyzed in the presence of additive white Gaussian noise (AWGN) channel. First we analyze the BER performance of an I-ary PAM scheme. Regular patterns in the k-th bit error probability are observed while developing the EBR expression. From these patterns we provide the exact and general closed-from EBR expression of an I-ary PAM. Then we present a general closed-from expression for BER of an arbitrary IXJ rectangular QAM by considering that this signaling format consists of two PAM scheme, i.e., I-ary and J-ary PAM. A simple approximate BER expression for rectangular QAM is given as well.