• Title/Summary/Keyword: mRNA amount

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Interrelationship between Cell Differentiation and Expression of mRNA for Transferrin in HL-60 Leukemia Cell Line

  • Lee, Soo-Young;Chi, Chung-Hee;Kim, You-Mie
    • BMB Reports
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    • v.33 no.4
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    • pp.308-311
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    • 2000
  • The interrelationship between the differentiation and expression of mRNA for transferrin in the HL-60 leukemia cell line was studied. Transferrin mRNA was expressed in HL-60 leukemia cells and the amount was 50% of that in the positive control cell line, HepG-2 cells. The expression of $T_f$ mRNA in HL-60 cells was not regulated by IL-1, IL-6 and $TNF-{\alpha}$, respectively. The expression of $T_f$ mRNA in the differentiated cells into a granulocyte lineage by DMSO, or all-trans RA, was up-regulated (160-170% of control cells); whereas, the expression was not regulated in the differentiated cells into a macrophage lineage by PMA. These results suggest that the differentiation to a granulocyte lineage of HL-60 leukemia cells appear to be related with the upregulation of transferrin mRNA expression.

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Effects of Alpha 1- and Alpha 2-Adrenoreceptor Stimulation on Galanin mRNA Expression in Primary Cultured Superior Cervical Ganglion Neurons

  • Xing, Yi;Chen, Xiuying;Liu, Zhen;Li, Hao;Liu, Huaxiang;Li, Zhenzhong
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.315-319
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    • 2011
  • Galanin (Gal) is a 29-amino-acid neuropeptide which is expressed in superior cervical ganglion (SCG) neurons and plays a trophic role in the adult animal and acts as an inhibitory modulator of cholinergic and noradrenergic neurotransmission. Whether activation or inhibition of alpha-adrenoreceptors infl uences Gal mRNA expression in SCG neurons remains unknown. Here, we have evaluated the possible regulation of Gal mRNA expression with acute (4 h) and chronic (4 days) stimulation of alpha 1- and alpha 2-adrenoreceptor agonists or antagonists in primary cultured SCG neurons. The results showed that the amount of Gal mRNA expression in cultured SCG neurons increased signifi cantly after chronic stimulation with alpha 2-adrenoreceptor antagonist yohimbine compared with control SCG neurons at the same time point, whereas the amount of Gal mRNA expression decreased signifi cantly after chronic stimulation with alpha 2-adrenoreceptor agonist clonidine as compared with that in control group. All these effects were not dose-dependent on the administration of alpha 2-adrenoreceptor agonist clonidine or alpha 2-adrenoreceptor antagonist yohimbine. Alpha 1-adrenoreceptor agonist phenylephrine or antagonist prazosin chronic stimulation did not have effects on Gal mRNA expression. Acute exposure of these agents did not have effects on Gal mRNA expression. The present study showed that Gal may be regulated by activation or inhibition of alpha 2-adrenoreceptors, but not alpha 1-adrenoreceptors in sympathetic neurons.

Isolation of High-Quality mRNA from Tannin-Rich Persimmon Fruit (고 Tannin 함유 감과실로 부터 mRNA의 분리)

  • ;Dav
    • Food Science and Preservation
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    • v.4 no.1
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    • pp.45-51
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    • 1997
  • In our studies on the role of $\beta$-galactosidase in fruit softening, significant difficulty, was encountered in our attempts to extract RNA from persimmon(Diospyros kaki L. cv. Fuyu) fruit due to astringency and tannin content. Initial, unsuccessful RNA extractions involved methods using guanidinium isothiocyanate/CsCl with and without polyvinylpyrrolidone(PVP), phenol/sodium lauryl sulfate(SDS), guanidinium hydrochloride, as well as polysomal RNA purification method that used 0.2 M Tris-HCI (pH 9.0) containing KCI, Mg-acetate, EDTA, $\beta$-mercaptoethanol, and sucrose. A method was devised which employed treatment of fruit with CO2 gas to diminish astringency prior to RNA extraction, followed by extraction of tissue powders with Proteinase K extraction buffer containing PVP and ascorbate at an alkaline pH. This procedure resulted in the removal of tannins and other polyphenolics and extraction of relatively large amount of high-quality RNA suitable for cDNA library construction and polymerase chain reaction(PCR). Futhermore, the procedure does not use the toxic and corrosive chemical guanidinium isothiocyanate or require ultracentrifugation.

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Expression of Taurine Transporter in Cell Lines and Murine Organs (세포주와 마우스 조직에서 타우린수송체의 발현분석)

  • 김하원;안희창;안혜숙;현진원;이은방
    • Biomolecules & Therapeutics
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    • v.10 no.2
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    • pp.78-84
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    • 2002
  • Taurine (2-ethaneaminosulfonic acid, $^+{NH}_3{CH_2}{CH_2}{SO_3^{-}}$) is endogenous amino acid with functions as modulator of osmoregulation, antioxidation, detoxification, transmembrane calcium transport, and a free radical scavenger in mammalian tissues. Taurine transporter(TAUT) contains 12 transmembrane helices, which are typical of the $Na^+$- and $Cl^-$-dependent transporter gene family, and has been cloned recently from several species and tissues. To analyze the expression of TAUT mRNA, one step RT-PCR was performed from human and mouse cultured cell lines and from various mouse tissues. The primers were designed to encode highly conserved amino acid sequences at the second transmembrane domain and at the fourth and fifth intracellular domains. RT-PCR analysis showed both of the human intestine HT-29 and mouse macrophage RAW264.7 cell lines expressed mRNA of TAUT. To define the expression patterns of the TAUT mRNA in the murine organs, RT-PCR was performed to detect cDNA representing TAUT mRNA from seven different mouse tissues. The TAUT was detected in all of the mouse tissues analyzed such as heart, lung, thymus, kidney, liver, spleen and brain. A large amount of transcript was fecund from heart, liver, spleen, kidney, and brain, while lung contained a very small amount of transcript.

Effects of $K^+$ lon on in vitro RNA Splicing of T4 Phage Thymidylate Synthase Gene

  • Sung, Jung-Suk;Park, In-Kook
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.49-53
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    • 1996
  • The effects of K$^{+}$ ion on the activity of RNA splicing of T4 phage thymidylate synthase gene have been investigated. The splicing activity was stimulated within the range of 5 to 20 mM concentration of KCI. When the concentration of KCI in the splicing reaction was brought to 100 or 200 mM a small amount of the exonl-intron product (1, 4 kb) was formed with large proportion of primary RNA transcript not undergoing splicing. This observation strongly suggests that there may exist come kinds of interferences with transesterification at the first step of splicing. Overall it can be concluded that K$^{+}$ ion exhibits very unique roles in RNA splicing of tdd gene depending on its concentration.ion.

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Effects of Overexpression of C5 Protein on rnpB Gene Expression in Escherichia coli

  • Kim, Yool;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.4
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    • pp.791-793
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    • 2009
  • Escherichia coli RNase P is composed of a large RNA subunit (M1 RNA) and a small protein subunit (C5 protein). Since both subunits are assembled in a 1:1 ratio, expression of M1 RNA and C5 protein should be coordinately regulated for RNase P to be efficiently synthesized in the cell. However, it is not known yet how the coordination occurs. In this study, we investigated how overexpression of C5 protein affects expression of the rnpB gene encoding M1 RNA, using a lysogenic strain, which carries an rnpB-lacZ transcription fusion. Primer extension analysis of rnpB-lacZ fusion transcripts showed that the overexpression of C5 protein increased the amount of the fusion transcripts, suggesting that rnpB expression increases with the increase of intracellular level of C5 protein.

The Analysis of Expression of Autoinducer Synthesis Genes Involved in Quorum Sensing among Catheter Associated Bacteria (요로감염에 관여하는 카테터 내 박테리아의 Quorum Sensing 관련 autoinducer 합성 유전자의 발현분석)

  • Lee, Mi-Hye;Seo, Pil-Soo;Lee, Ji-Youl;Peck, Kyong-Ran;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.277-285
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    • 2006
  • The most biofilm forming bacteria in catheter, Esctherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were isolated and identified from a patient's catheter occuring catheter-associated urinary tract infection (CA-UTI). We examined mRNA expression and its quantification of AIs synthetic genes encoding signal substance of quorum sensing from each bacterial species in order to elucidated quorum sensing mechanism. Both pure cultures for each bacterial strains and a mixed cultures with three were grown for 24 hr and 30 days. Initial densities to be able to detect mRNA expression oil single strains culture were shown at $2.4{\times}10^5$ CFU/ml, $5.4{\times}10^6$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $6.9{\times}10^4$ CFU/ml of P. aeruginosa for rhlI and lasI. Also, in mixed culture of three, initial cell densities of mRNA expression were appear to at $7.3{\times}10^5$ CFU/ml, $1.6{\times}10^7$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $2.1{\times}10^5$ CFU/ml of P. aeruginosa for rhlI and lasI. Each AIs synthetic gene was expressed in initial cell density and the mRNA expression of the genes were detected continously during 30 days. And then, the quantification of mRNA expression level of ygaG, rhlI, last, and luxS which were related AIs synthesis was done each time point by real-time RT-PCR. Interestingly, the mRNA levels of ygaG, rhlI, lasI, and luxS from the mixed culture was higher than those from each single strain culture. In the case of E. coli ygaG, the amount of transcript from the mixed culture was at least 30 times for that from single culture. In the case of P. aeruginosa rhlI and lasI, the amount of transcript from the mixed culture was at least 40 times and 250 times for that from single strain culture. In the case of S. aureus luxS, the amount of transcript from the mixed culture was at least 5 times for that from single strain culture. And specially, the mRNA expression of rhlI and lasI of P. aeruginosa showed the highest efficency among four AIs synthetic genes.

Antioxidant and Whitening Activities of Chlorogenic Acid, Quercetin, and Quercitrin from the Fruit of Vaccinum oldhami (정금나무 열매(Fruit of Vaccinum oldhami)의 분리 정제물(클로로겐산, 퀘르세틴 및 퀘르시트린)에 관한 항산화 및 미백활성 검증)

  • Jung-Woo Chae;Min-Jeong Oh;Hyeon-Ji Yeom;Jin-Young Lee
    • Journal of Life Science
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    • v.33 no.2
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    • pp.115-128
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    • 2023
  • The fruit of Vaccinum oldhami was separated and purified to obtain the compounds chlorogenic acid (CA), quercetin (QT), and quercitrin (QR). The electron-donating abilities of CA, QT, and QR at 1,000 ㎍/ml were 91.9%, 89.9%, and 77.4%, respectively QT and QR showed 99.5% and 91.4% ABTS+ radical scavenging ability at a 1,000 ㎍/ml concentration, respectively, and CA showed a 95% ability or higher at 100 ㎍/ml. Regarding tyrosinase inhibitory activity, CA, QT, and QR exhibited 29.5%, 34.7%, and 23.7% efficacy, respectively, at 1,000 ㎍/ml. Regarding the cell viability for melanoma cells (B16F10) assessed through MTT assay, CA, QT, and QR showed cell a viability of 80% or more at 100 ㎍/ml. To measure the deterrent of protein expression, CA affected TRP-1 and TRP-2 in accordance with increases in concentration. The protein expression inhibition rate of QT was excellent for TRP-1, TRP-2, and tyrosinase. CA was confirmed to have an excellent mRNA expression inhibitory effect against MITF, and the amount of mRNA expression of TRP-1, TRP-2, and tyrosinase decreased with an increase in the CA concentration. As the concentration of QT increased, the mRNA expression of MITF, TRP-2, and tyrosinase decreased. QR decreased the amount of mRNA as the QR concentration increased. The excellent antioxidant and whitening effects of CA, QT, and QR were thus confirmed.

Cold Shock Response and Low Temperature Stable Transcript of DEAD-box RNA Helicase in Bacillus subtilis (DEAD-box RNA Helicase 유전자가 결핍된 Bacillus subtilis의 저온 충격 반응성과 저온 안정성 전사물)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.289-294
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    • 2011
  • We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

Effects of Acarbose on the Expression of Obese and Neuropeptide Y (NPY) Genes in Mice on High-Carbohydrate Diet

  • Kim, Ji-Yeon;Chung, Sung-Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.4
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    • pp.433-438
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    • 1999
  • Two components of the neuroendocrine-hormonal response to long-term treatment of acarbose, adipose tissue-derived leptin and central neuropeptide Y (NPY), were investigated in the ICR mice on a high- carbohydrate diet. Acarbose, administered 5 or 50 mg per 100 g diet for four weeks, dose dependently suppressed body weight gain. The body weight gain was reduced along with the amount of daily food intake in 50 mg acarbose-treated group at $7^{th}\;and\;28^{th}$ day. 5 or 50 mg acarbose treatment administered for four weeks reduced leptin mRNA levels to 62% and 77% of the control group, demonstrating that the amount of leptin mRNA in adipocytes correlates with body weight. As dose of acarbose increased, leptin mRNA level also increased, suggesting that potent inhibition of ${\alpha}-glycosidase$ by a higher dose of acarbose furthers the enzyme activity and leptin gene consequently. On the other hand, central expression level of NPY gene was increased significantly compared with the control group at the same amount of acarbose administered, reflecting that leptin and NPY operate in a negative-feedback circuit to regulate body fat stores.

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