• Title/Summary/Keyword: RNA content

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Considering Cell-based Assays and Factors for Genome-wide High-content Functional Screening

  • Chung, Chul-Woong;Kim, In-Ki;Jung, Yong-Keun
    • Animal cells and systems
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    • v.13 no.2
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    • pp.97-103
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    • 2009
  • Recently, great advance is achieved in the field of genome-wide functional screening using cell-based assay. Here, we briefly introduce well-established and typical cell-based assays of GPCR and some parameters which should be considered for genome-wide functional screening. Because of characters and importance of GPCR as drug targets, several ways of assay systems were devised. Among them, high-content screening (HCS) that is based on the analysis of image by confocal microscope is becoming favorite choice. The advances in this technology have been driven exclusively by industry for their convenience. Now, it is turn for academy to define more detail signaling networks via HCS using cDNA or siRNA libraries at genome-wide level. By isolating novel signaling mediators using cDNA or siRNA library, and postulating them as new candidates for therapeutic target, more understanding about life science and more increased chances to develop therapeutics against human disease will be achieved.

Correlation between Heart-type Fatty Acid-binding Protein Gene Polymorphism and mRNA Expression with Intramuscular Fat in Baicheng-oil Chicken

  • Wang, Yong;He, Jianzhong;Yang, Wenxuan;Muhantay, Gemenggul;Chen, Ying;Xing, Jinming;Liu, Jianzhu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1380-1387
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    • 2015
  • This study aims to determine the polymorphism and mRNA expression pattern of the heart-type fatty acid-binding protein (H-FABP) gene and their association with intramuscular fat (IMF) content in the breast and leg muscles of Baicheng oil chicken (BOC). A total of 720 chickens, including 240 black Baicheng oil chicken (BBOC), 240 silky Baicheng oil chicken (SBOC), and 240 white Baicheng oil chicken (WBOC) were raised. Three genotypes of H-FABP gene second extron following AA, AB, and BB were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) strategy. The G939A site created AA genotype and G956A site created BB genotype. The content of IMF in AA genotype in breast muscle of BBOC was significantly higher than that of AB (p = 0.0176) and the genotype in leg muscle of WBOC was significantly higher than that of AB (p = 0.0145). The G939A site could be taken as genetic marker for higher IMF content selecting for breast muscle of BBOC and leg muscle of WBOC. The relative mRNA expression of H-FABP was measured by real-time PCR at 30, 60, 90, and 120 d. The IMF content significantly increased with age in both muscles. The mRNA expression level of H-FABP significantly decreased with age in both muscles of the three types of chickens. Moreover, a significant negative correlation between H-FABP abundance and IMF content in the leg muscles of WBOC (p = 0.035) was observed. The mRNA expression of H-FABP negatively correlated with the IMF content in both breast and leg muscles of BOC sat slaughter time.

Development of Saccharomyces cerevisiae Strains with High RNA Content (리보핵산을 다량으로 함유하는 Saccharomyces cerevisiae 균주의 개발)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Min, Byoung-Cheol;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.465-474
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    • 1999
  • RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

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Nutritional and Hormonal Regulation of Fatty Acid Synthase Gene Expression

  • Shin, Dong-Hoon;Kim, Byung-Yong;Hahm, Young-Tae;Kim, Eunki;Cho, Won-Dai
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.246-249
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    • 1997
  • The maximum 30-fold level of fatty acid synthase (FAS) mRNA was achieved by 6hr after intraperitoneal injection of insulin. The kinetics and maximum effect of insulin were most evident on he 7.2 kb mRNA. In six hors after insulin administration there was about 100-fold increase in stead-state mRNA level. We observed a sharp decrease in 7.2kb mRNA by 8hr after insulin administation while there was no change in FAS mRNA content between the 6hr and 8hr-sampling periods. In contrast, a maximum induction of 4-fold was shown in the level of 5.1kb mRNA after insulin injection in streptozotocin-diabetic mice.

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Daidzein Modulations of Apolipoprotein B and Fatty Acid Synthase mRNA Expression in Chick Liver Vary Depending on Dietary Protein Levels

  • Choi, Jinho;Song, Jungmin;Choi, Yeon-Mi;Jang, Dong-Ju;Kim, Eunmi;Kim, Inho;Chee, Kew-Mahn
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.236-244
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    • 2006
  • This study was designed to determine the effects of daidzein (DE) on hepatic lipid metabolism in chicks fed with low protein (LP) diet based on casein. In experiment 1, the male chicks were fed with one of the three levels of dietary protein containing 10.95%, 21.9% and 43.8% protein content for 2 days. In experiment 2, the chicks were fed one of the three levels of protein with or without DE at 1,000 mg/kg diet for 2 days. Experiment 3 was conducted to compare DE (LP+DE) with estradiol (LP+E2) in chicks fed with LP diet for 7 days. Plasma lipid profiles, hepatic lipid profiles, activities of hepatic malic enzyme and isocitrate dehydrogenase (ICDH) were measured. Transcriptions of hepatic fatty acid synthase, apolipoprotein-B (APO-B), and fructose bisphosphatase mRNA were measured by RT-PCR. Increasing dietary protein levels markedly decreased the concentrations of plasma triglycerides, hepatic total lipids, hepatic TG, and the mRNA transcriptions while the increased dietary protein levels increased hepatic ICDH activities in experiment 1. In experiment 2, the effects of dietary protein levels on blood and hepatic lipid content were more prominent than those of the additional DE. Interestingly, plasma TG levels were affected by DE supplementation (p<0.05). In experiment 3, DE inhibited APO-B mRNA expressions and stimulated the accumulation of lipid in the liver through mechanisms different from E2. In this study, we demonstrate that DE has beneficial effects on blood lipid profiles, but that it inhibits APO-B mRNA transcription and aggravates the fatty liver induced by LP diet in chicks.

Complete genome sequence of Bacillus thuringiensis C25, a potential biocontrol agent for sclerotia-forming fungal phytopathogens (생물학적방제 효과가 뛰어난 Bacillus thuringiensis C25 균주의 유전체 분석)

  • Lee, Hwa-Yong;Won, Kyungho;Kim, Yoon-Kyeong;Cho, Min;Kim, Kangmin;Ryu, Hojin
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.216-218
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    • 2017
  • We here provide the complete genome sequence of Bacillus thuringiensis C25, the strain showing antagonistic effects on fungal phytopathogens. The genome comprised of 5,308,062 bp with 35.32% G+C content of a circular chromosome and a plasmid containing 308,946 bp with 32.23% G+C content. The chromosome and plasmid genome included 5,683 protein coding DNA sequences, 107 tRNA and 42 rRNA genes.

Biochemical Studies of Ginseng Saponin on RNA and Protein Biosynthesis in the Rat Liver (간에서의 RNA, 단백질 생합성에 미치는 인삼성분의 생화학적 연구)

  • Oura Hikokichi
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.1-10
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    • 1988
  • Previously. we reported that the intraperitoneal administration of ginseng crude saponin increased: (I) nuclear RNA polymerase activity. (2) nuclear RNA synthesis. (3) cytoplasmic RNA synthesis. (4) cytoplasmic heavy polyrioosome content. (5) amino acid incorporation in vitro of microsome and polysome isolated rat liver. and (6) the incorporation rate of labeled amino acids into serum protein. In addition, a spectacular increase in the rough endoplasmic reticulum of hepatocyte administered crude saponin for four weeks orally was shown through electron microscopy. An increase in polysomal content in membrane-hound ribosome was shown through ultracentrifugation. Recently, successive intraperitoneal. administration .of $ginsenosid-Rb_2$ was given to streptozotocin (STZ) diaoetic rats of hypoproteinemia. The blood urea nitrogen and hepatic urea concentration were decreased significantly. The total protein and alhumin levels in the serum were increased in comparison to control values. In contrast. the $ginsenoside-Rb_2$ treated group of STZ diahetic rats showed a significant increase in liver RNA. total ribosome and membrane-bound ribosomal contents. The administration of $ginsenoside-Rb_2$ increased the incorporation rate of labeled - precursor into total serum protein. Additionally $ginsenoside-Rb_2$ improved the nitrogen balance of diabetic rats. On the bases of these experimental results, ginseng saponin has a metabolic stimulatory or anabolic action on RNA and protein synthesis.

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Studies on Physicochemical and Biological Properties of Depolymerized Alginate from Sea tangle, Laminaria japonicus by Thermal Decomposition 5. Effects of Depolymerized Alginate on Body Weight, Organ, Pancreatic and Small Intestinal Composition, and Small Intestinal Microvilli Structure in Rats (다시마 (Laminaria japonicus) Alginate의 가열가수분해에 따른 물리$\cdot$화학적 및 생물학적 특성에 관한 연구 5. 랫드의 체중, 장기, 췌장과 소장의 성분 및 소장융모의 미세구조에 미치는 저분자 Alginate의 영향)

  • KIM Yuck-Yong;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.1
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    • pp.21-26
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    • 2001
  • To examine functionality of depolymerized alginate obtained by hydrolysis of alginate through a heating process at $121^{\circ}C$ on gastrointestinal physiology, the changes of body weight, organ weight and length, pancreatic and small intestinal composition, and light microscopy (LM) observation of small intestinal microvilli's appearances were checked in the rats. Rats were fed diets containing $1\%, 5\%, and 10\%$ of each depolymerized alginate (HAG-10, HAG-50, HAG-100) and alginate for 35 days, The feeding of 5 and $10\%$ HAG-50 and $10\%$ alginate diets for 35 days significantly depressed the body weight gain, but increased the length and weight of the small intestine and cecum in rats (p<0.01). Pancreatic protease activity was decreased significantly (p<0.01) in all groups except lo/o of HAG-10 diets, but the protein content increased in all groups, However, pancreatic amylase and lipase activities as well as DNA and RNA content were not significantly different. The small intestinal protein and the DNA content were the highest in diets fed $5\%$ HAG-50; RNA content increased significantly (p<0.01) in all groups except in the fiber-free diets. Light microscopy (LM) observation showed growth of small intestinal microvilli with numerous ridges; the multiplication of the convolution goblet cells in rats fed with diets containing $5\%$ of HAG-50 were more than others group.

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Response of Monodehydroascorbate Reductase in Lettuce Leaves Subjected to Low Temperature Stress (저온 처리한 상추 잎에서 monodehydroascorbate 환원효소의 반응)

  • Kang, Sang-Jae
    • Journal of Life Science
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    • v.21 no.3
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    • pp.368-374
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    • 2011
  • The relationship between MDHA reductase activity and ascorbate, dehydroascorbate, and hydrogen peroxide content was evaluated, and this experiment was conducted to determine the change of MDHA reductase activity and the level of steady-state mRNA abundance of MDHA reductase in lettuce leaves subjected to low temperature stress. MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ for 24 hr increased, followed by a steady decrease during the duration of recovery to $20^{\circ}C$ for 48 hr. The content of ascorbate slowly increased during low temperature treatment, followed by a rapid increase during the duration of recovery to $20^{\circ}C$ for 48 hr, while dehydroascorbate content rapidly decreased. The relationship between MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ and ascorbate content correlated positively ($R^2$=0.9240, 0.9108, respectively), but MDHA reductase activity of chloroplastic and cytosolic fraction and dehydroascorbate were reversely correlated ($R^2$=0.8638, 0.8980, respectively). Hydrogen peroxide content and MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ correlated positively ($R^2$=0.9443, 0.9647, respectively). Northern blot analysis showed that the level of mRNA transcript of MDHA reductase was similar to total activity of MDHA reductase, and also that the level of mRNA of MDHA reductase after recovery to $20^{\circ}C$ for 24 hr decreased.

Modification of Tomato Aspermy Virus Symptom by Cucumber Mosaic Virus- Associated Satellite RNA (Cucumber Mosaic Virus-associated Satellite RNA에 의한 Tomato Aspermy Virus의 병징변화)

  • Lee Hyun Sook
    • Korean Journal Plant Pathology
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    • v.2 no.3
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    • pp.145-149
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    • 1986
  • A tomato Aspermy Virus (TAV-B) served as a helper virus for multiplication and encapsidation of satellite RNAs which were isolated from two different CMV isolates, D and K. These two satellite RNAs induced renarkable attenuation of TAV symptoms in infected tobacco, which was correlated with a reduction of virus content in the plant. The CMV satellite RNAs also caused lethal necrosis in TAV-infected tomato as in the case of CMV system.

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