• Title/Summary/Keyword: DNA Synthesis

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Differential Regulation of the Genes of the Streptococcus pneumoniae dnaK Operon by Ca++

  • Kim, Seung-Whan;Bae, Yong-Goo;Pyo, Suhk-Neung;Rhee, Dong-Kwon
    • Molecules and Cells
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    • v.23 no.2
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    • pp.239-245
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    • 2007
  • DnaK is a major antigen in Streptococcus pneumoniae, and is induced by a minor shift in temperature (30 to $37^{\circ}C$) but not by ethanol shock. Although HrcA in the presence of $Ca^{{+}{+}}$ represses the expression of both groEL and hrcA, the control of transcription of the dnaK operon is not completely understood. In this study, the dnaK operon of S. pneumoniae (5' hrcA-grpE-dnaK-dnaJ) was cloned and analyzed. It contains large intergenic regions in grpE/dnaK and dnaK/dnaJ. Pulse labeling with [$^{35}S$]-methionine and immunoblot analyses revealed the presence of higher levels of DnaK than of HrcA even in the presence of $Ca^{{+}{+}}$ after heat shock suggesting that $Ca^{{+}{+}}$ differentially regulates the heat shock responses of hrcA and dnaK. By blocking de novo mRNA synthesis with rifampin it was shown that neither the hrcA nor the groEL transcripts were stabilized by heat shock even though dnaK transcripts were stabilized. We conclude that S. pneumoniae uses fine regulation of the transcription of the individual genes of the tetracistronic dnaK operon to cope with the various stresses experienced during infections.

Synthesis and characterization of doxorubicin hydrochloride drug molecule-intercalated DNA nanostructures

  • Gnapareddy, Bramaramba;Deore, Pragati Madhukar;Dugasani, Sreekantha Reddy;Kim, Seungjae;Park, Sung Ha
    • Current Applied Physics
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    • v.18 no.11
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    • pp.1294-1299
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    • 2018
  • In this paper, we demonstrate the feasibility of constructing DNA nanostructures (i.e. DNA rings and double-crossover (DX) DNA lattices) with appropriate doxorubicin hydrochloride (DOX) concentration and reveal significant characteristics for specific applications, especially in the fields of biophysics, biochemistry and medicine. DOX-intercalated DNA rings and DX DNA lattices are fabricated on a given substrate using the substrateassisted growth method. For both DNA rings and DX DNA lattices, phase transitions from crystalline to amorphous, observed using atomic force microscopy (AFM) occurred above a certain concentration of DOX (at a critical concentration of DOX, $30{\mu}M$ of $[DOX]_C$) at a fixed DNA concentration. Additionally, the coverage percentage of DNA nanostructures on a given substrate is discussed in order to understand the crystal growth mechanism during the course of annealing. Lastly, we address the significance of optical absorption and photoluminescence characteristics for determining the appropriate DOX binding to DNA molecules and the energy transfer between DOX and DNA, respectively. Both measurements provide evidence of DOX doping and $[DOX]_C$ in DNA nanostructures.

Effect of Sinhyo-Taklee-San on the Proliferation of Fibroblast Cell(Balb/c 3T3) (섬유아세포(纖維芽細胞)(Balb/c 3T3)의 증식(增殖)에 미치는 신효탁리산(神效托裡散)의 영향(影響))

  • Eun, Jae-Soon;Jeon, Young-Keun;Yum, Jeong -Yul;Suh, Eun-Sil;So, June-No;Oh, Chan- Ho
    • Korean Journal of Pharmacognosy
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    • v.24 no.2
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    • pp.159-165
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    • 1993
  • The studies were conducted to investigate the effect of Sinhyo-Taklee-San(STS), which is composed of Astragali Radix(AR), Lonicerae Flos(LF), Angelicae gigantis Radix(AGR) and Glycyrrhizae Radix(GR), on the proliferation of fibroblast cell(Balb/c 3T3). STS, GR and glycyrrhizin increased the proliferation of 3T3 cells. The 10% serum obtained from STS, AR, LF, AGR and GR treated mice also increased the proliferation of 3T3 cells markedly. GR, glycyrrhizin and glycyrrhetinic acid inhibited protein synthesis, but did not affect on DNA synthesis.

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A gene responsible for ozone sensitivity (ozrB) in chromosome of escherichia coli B. MQ 1844 (E. coli B. MQ1844 균주의 오존감수성 유전자의 특성)

  • ;Hamelin C.
    • Korean Journal of Microbiology
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    • v.25 no.2
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    • pp.103-109
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    • 1987
  • An ozone-sensitive mutant of Escherichia coli strain B, MQ 1844 is described. Its properties, including high sensitivity to ozone and radiation, inducible filamentation, extensive DNA degradation and impaired DNA synthesis following ozonation, are attributable to a mutation in ozrB, a gene which is cotransducible with malB. Based on differences in phenotypic expression as well as on the particular location of this gene on the bacterial chromosome, ozrB appears as distinct from the other ozone-or radiation-sensitivity genes previously described.

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Development of DNA Markers for Trehalose Synthesis Genes in Brassica rapa L. (배추 trehalose 합성 유전자와 연관된 DNA 마커 개발)

  • Jeong, Ye-Sol;Lim, Yong-Pyo;Hur, Yoon-kang;Chung, Sang-Min
    • Journal of Life Science
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    • v.19 no.5
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    • pp.639-643
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    • 2009
  • High temperature stress might affect the yield and quality of Chinese cabbage. In order to develop cultivars resistant to high temperature stress, we developed polymorphic DNA markers for trehalose synthesis genes related to abiotic stress resistance. A total of 28 Brassica rapa ESTs homologous to trehalose synthesis genes of Arabidopsis were found from the NCBI database. The polymorphic DNA sequences were searched between Chinese cabbages - Chiifu, which is relatively susceptible to high temperature stress, and Kenshin, which is tolerant to high temperature stress. Among the 28 ESTs, we found 10 ESTs that have either insertion/deletion and/or single nucleotide polymorphism between the two cultivars. Those polymorphic sites were then targeted for the development of 10 PCR based markers. These molecular markers related to trehalose genes could be used not only to test their relationship with abiotic stress resistance in Chinese cabbage, but also the development of abiotic stress resistant cultivars using MAS.

Synthesis and base pairing properties of DNA-RNA heteroduplex containing 5-hydroxyuridine

  • Cui, Song;Kim, Yong-Hoon;Jin, Cheng-Hao;Kim, Sang-Kook;Rhee, Man-hee;Kwon, Oh-Shin;Moon, Byung-Jo
    • BMB Reports
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    • v.42 no.6
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    • pp.373-379
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    • 2009
  • 5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-ODMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-$Script^{TM}II$-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU $\approx$ C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5- OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5- OHU from oxidative damage was mainly influenced by adenine mismatch.

Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
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    • v.24 no.2
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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Effects of Gamgung-tang on Proliferation and cAMP Accumulation of Thyroid Cells (감궁탕이 갑상선세포의 증식과 cAMP 축적에 미치는 영향)

  • Kim, Mee-Kyung;Shon, Yun-Hee;Nam, Kyung-Soo;Son, Ok-Lye;Kim, Cheorl-Ho;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.623-627
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    • 2005
  • Abnormal thyroid cell proliferation has a very important role in hypothyroidism. Thyroid stimulating hormone (TSH) stimulates proliferation and maintains differentiated function in thyroid follicular cells. A functioning rat thyroid cell line (FRTL) was used to study the effect of Gamgung-tang (GGT, Glycyrrhizae Radix, black beans, Angelicae Radix and Cnidii Rhizoma) on proliferation and cAMP accumulation of thyrocytes. Proliferation of cell was assessed by DNA synthesis and incorporation of $[^3H]thymidine$ into DNA. The concentration of cAMP was measured simultaneously with growth assessment. Extract of GGT ($0.15{\sim}0.9\;mg/ml$ increased DNA synthesis in a dose-dependent manner. GGT at 0.6 (p<0.05) and 0.9 mg/ml (p<0.01) significantly increased $[^3H]thymidine$ incorporation. A comparable effect was observed with TSH. GGT also enhanced cAMP accumulation. These results indicate that GGT increases the proliferation of thyrocytes and may be considered a promising agent for the treatment of autoimmune thyroid disease.

Growth Inhibitory Effects of Sesamolin from Sesame Seeds on Human Leukemia HL-60 Cells (참깨에서 분리된 세사몰린의 백혈병 세포주 HL-60 생장억제 효과)

  • Kim, Kwan-Su;Kang, Sam-Sik;Ryu, Su-Noh
    • Korean Journal of Pharmacognosy
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    • v.34 no.3 s.134
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    • pp.237-241
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    • 2003
  • This study was carried out to test the growth inhibitory effects of sesamolin obtained from sesame seeds. Sesamolin inhibited the growth of human leukemia HL-60 cells in cultures and the synthesis of macromolecules in dose- and time-dependent manners. Sesamolin in the $60{\sims}100\;{\mu}g/ml$ range was cytostatic. At concentrations greater than $200\;{\mu}g/ml$ sesamolin was cytocidal to HL-60 cells and at $60\;{\mu}g/ml$ inhibited the synthesis of DNA, RNA and protein in HL-60 cells by 35.1, 6.1, and 5.3%, whereas at $200\;{\mu}g/ml$ these inhibitions were 86.8%, 81.5% and 96.7%, respectively. The inhibitory effect of sesamolin on DNA synthesis was irreversible.