• Title/Summary/Keyword: RNA/DNA

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Effect of Several Drugs of DNA, RNA and Protein Damage induced by Dimethylnitrosamine in Mouse Tissues (수종약물이 Dimethylnitrosamine에 의한 DNA, RNA 및 단백질 손상도에 미치는 영향)

  • Kim, Jea-Hyun;Park, Jung-Sik;Hong, Sung-Ryul;Kweon, O-Cheul;Park, Chang-Won;Rhee, Dong-Kwon
    • YAKHAK HOEJI
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    • v.35 no.6
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    • pp.522-529
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    • 1991
  • The purpose of this research is to evaluate effects of chloramphenicol, phenobarbital and progesterone on damage of DNA, RNA and protein which was induced by dimethylnitrosamine. $N,N-Di[^{14}C]$ methyl-nitrosamine (DMN) was used as a damaging agent and levels of DNA, RNA and protein damage in liver, brain and pancreas were compared with a control group. Pretreatment of mice with chloramphenicol increased protein damage in pancreas two times more than the control level. Liver RNA damage was increased up to 5.8 times and brain DNA damage up to 6.95 times by treatment of phenobarbital but brain RNA damage was decreased significantly down to 21% of the control group. The damage of liver RNA was significantly decreased by treatment of progesterone, although liver protein damage, pancreas RNA damage and pancreas protein damage were increased.

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Characterization of the Folding Structure of 3'-end of Lactate Dehydrogenase A-mRNA Isolated from Hormone Stimulated Rat $C_{6}$ Glioma cell culture (홀몬으로 처리된 쥐의 $C_{6}$ glioma 세포배양으로부터 분리된 낙산탈수소 효소 A-mRNA의 3'-말단의 2차 구조)

  • 배석철;이승기
    • Korean Journal of Microbiology
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    • v.25 no.2
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    • pp.94-102
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    • 1987
  • Rat liver LDH A-cDNA has been isolated from a .lambda.gt11-rat lover cDNA library and partially characterized. The size of the isolated rat liver LDH A-cDNA if shown to be 1.6Kb and restriction enzyme sites for the rat liver LDH A-cDNA are also mapped. 682-nucleotide sequence coding for 3'-end of rat liver LDH A-cDNA has been analyzed and compared to the nucleotide sequence of the same region of rat $C_6$-glioma cell LDH A-cDNA which has been cloned from the hormonally stimulated cell cultures. The result shows that 177 nucleotide sequences coding for the C-terminal 59-amino acids are identical but 505 nucleotide sequences of 3'-nontranslated region of the two LSH A-cDNA exhibit characteristic differences in thier nucleotide sequences. Computer analysis for the folding structures for 3'-end 400 nucleotide sequences of the two LDH A-cDNA shows a possibility implying that the two LDH A-mRNAs isolated from different tissues of rats may have different half life and therefore their translational efficiency may be different. It has been previously demonstrated that isoproterenol stimulated rat $C_6$ -glioma cell cultures produce LDH A-mRNA showing 2 to 3-fold longer half life in comparison to that of noninduced LHD A-mRNA. The result therefore support for the idea that hormonally stimulated rat $C_6$-glioma cells may produce LDH A-mRNA containing different nucleotide sequences at the 3'-end nontranslated region by which the hormonally induced LDH A-mRNA could have more stable secondary mRAN structure in comparison to that of noninduced LDH A-mRNA.

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Construction of nervous necrosis virus (NNV) genome-based DNA replicon vectors for the delivery of foreign antigens

  • Jeong In Yang;Ki Hong Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.1-8
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    • 2024
  • The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate immunity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glycoprotein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glycoprotein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.

DNA, RNA, Protein and Yield of the Soybean Plant, Glycine max Merr., as Affected by Phosphorus Nutrition (대두의 핵산, 단백질 및 물질생산에 미치는 인산비료의 효과)

  • 장남기
    • Journal of Plant Biology
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    • v.16 no.1_2
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    • pp.23-29
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    • 1973
  • The effect of phosphorus nutrition on the content of deoxyribonucleic acid(DNA), ribonucleic acid(RNA), crude protein and plant growth of soybean plant(Glycine max, Merr.) was studied. Yields of the above-and under-ground parts of the soybean plant in terms of dry weight, the amounts of crude protein, RNA and DNA continued to increase with increasing phosphorus supply. The amounts of RNA and crude protein were highest in the leaf tissues where most intensive growth was taking place. The relationships among DNA, RNA, crude protein and plant growth appeared to consist of the central dogma which has immortalized, while DNA in plant tissue was subject to charges cuased by external environmental facters such as phosphorus nutrition.

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Seasonal Change of RNA/DNA Ratio and Blood Characteristics of Black Sea Bream Acanthopagrus schlegeli Habituated in Geojae Costal Area, Kyungnam Province, Korea (경남 거제해역에 서식하는 감성돔(Acanthopagrus schlegeli)의 연간 RNA/DNA 및 혈액학적 특성 변화)

  • Kim, Su-Kyoung;Shim, Na-Young;Lee, Do-Hyun;Kim, Dae Hyun;Yoon, Seong-Jong
    • Journal of Environmental Science International
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    • v.22 no.1
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    • pp.37-45
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    • 2013
  • The monthly variations of blood characteristics and RNA/DNA of black sea bream, Acanthopagrus schlegeli, habituated in Geojae costal area were analysed to determine health condition of natural stocks in terms of gonad maturation and spawning season from March 2010 to February 2011. Spawning season determinated by gonadosomatic index is from June to August. RNA/DNA ratio of black sea bream muscle was strongly correlated with spawning season. During the gonad maturation RNA/DNA ratio in dorsal muscle tissue was decreased contrast to rapid increase during spawning season. Blood composition factors increased in terms of gonad maturation are aspartate aminotransferase, cholesterol, triglyceride, total protein, glucose, globulin, alkaline phosphatase and inorganic phosphate. Other blood factors increased during spawning season are alanine aminotransferase, blood urea nitrogen, uric acid and lactate dehydrogenase.

Influence of Ginseng upon Testicular Nucleic Acid Content in Rats (인삼이 흰쥐의 고환조직 핵산 함유량에 미치는 영향)

  • Suh, Chung-Min;Kim, Byung-Ho;Chang, Im-Soo
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.37-40
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    • 1973
  • As a continuation of efforts to elucidate the influence of ginseng upon nucleic acid content of various tissues, a study was carried out which measured testicular RNA and DNA contents of rats following administration of ginseng. Thirty male rats $(body\;weight:\;180{\sim}230\;gm)$ were equally divided into a ginseng and a saline group. Once a day for 5 days they received subcutaneously 0.5 m1/100 gm body weight of ginseng extract solution (4 mg of ginseng alcohol extract in 1 ml of saline), and the same amount of satire, respectively. On the 5th experimental day, all animals were sacrificed 2 hours after the last medication and their testicular RNA and DNA contents were measured using the chemical method of Schmidt-Thannhauser-Schneider. Following results were obtained: 1. Testicular RNA and DNA contents were significantly higher in the ginseng group than in the saline group. 2. RNA/DNA ratio of the testis was lower in the ginseng group than in the saline group. However, the two groups did not differ significantly with regard to the RNA/DNA ratio. The ginseng is inferred to raise RNA and DNA contents of testis in rats.

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Ecophysiological Characteristics Changes in the Pacific Oyster, Crassostrea gigas, after Spawning Season in Off-bottom Culture (수평망식 양식 굴, Crassostrea gigas의 산란기 이후 생태·생리학적 특성 변화)

  • Kim, Su Kyoung;Choi, Eun Hee;Han, Hyun Seob;Lim, Hyun Jeong
    • The Korean Journal of Malacology
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    • v.28 no.3
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    • pp.215-223
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    • 2012
  • The ecophysiological characteristics of the oyster Crassostrea gigas in Taean area, west coast of Korea, were investigated after main spawning season from September 2011 to January 2012 to confirm the recovery process of condition. The cupped oysters, from 4 different off-bottom culture areas were compared the gonad indices, histological analysis combined with measurement of RNA/DNA contents and ratios in gonad of each sex were undertaken. To evaluate the health condition of oyster after spawning, the RNA/DNA ratio in adductor muscle, fatness and condition index (CI) were used. The results showed that cupped oysters cultured in Uihang coastal area were different physiological aspects than other experiment areas, namely continuously decreasing RNA/DNA content and ratio in adductor muscle, lowest CI and fatness. Oysters cultured in Iwon Station 1 and 2 showed fast recovery in RNA/DNA, increase in fatness and CI during post-spawning stage until October rapidly and there after it decreased. Oysters in Shinduri showed rapidly decrease in fatness and CI until October. On the contrary to this factors, RNA/DNA ratio in adductor muscle increased as like protein until October. Partially spawning season could be observed until December in Shinduri and Iwon Station 1.

Development and Validation of Quick and Accurate Cephalopods Grouping System in Fishery Products by Real-time Quantitative PCR Based on Mitochondrial DNA (두족류의 진위 판별을 위한 Real-time Quantitative PCR 검사법 개발 및 검증)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji Young;Kwon, Ki sung;Kim, Gun Do
    • Journal of Food Hygiene and Safety
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    • v.33 no.4
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    • pp.280-288
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    • 2018
  • In this study, an approach for the analysis of the five cephalopod species (octopus, long-arm octopus, squid, wet-foot octopus, beka squid) consumed in the Republic of Korea is developed. The samples were collected from the Southeast Asian countries Thailand, Indonesia, Vietnam, and China. The SYBR-green-based real-time qPCR method, based on the mitochondrial DNA genome of the five cephalopods was developed and validated. The intergroup variations in the mitochondrial DNA are evident in the bioinformatic analysis of the mitochondrial genomic DNA sequences of the five groups. Some of the highly-conserved and slightly-variated regions are identified in the mitochondrial cytochrome-c-oxidase subunit I (COI) gene, 16s ribosomal RNA (16s rRNA) gene, and 12s ribosomal RNA (12s rRNA) gene of these groups. To specify each five cephalopod groups, specific primer sets were designed from the COI, 16s rRNA and 12s rRNA regions. The specific primer sets amplified the DNA using the SYBR-green-based real-time PCR system and 11 commercially secured animal tissues: Octopus vulgaris, Octopus minor, Todarodes pacificus, Dosidicus gigas, Sepia esculenta, Amphioctopus fangsiao, Amphioctopus aegina, Amphioctopus marginatus, Loliolus beka, Loligo edulis, and Loligo chinensis. The results confirmed by a conveient way to calculate relative amplification levels between different samples in that it directly uses the threshold cycles (Ct)-value range generated by the qPCR system from these samples. This genomic DNA-based molecular technique provides a quick, accurate, and reliable method for the taxonomic classification of the animal tissues using the real-time qPCR.

Development of a Novel Long-Range 16S rRNA Universal Primer Set for Metagenomic Analysis of Gastrointestinal Microbiota in Newborn Infants

  • Ku, Hye-Jin;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.812-822
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    • 2014
  • Metagenomic analysis of the human intestinal microbiota has extended our understanding of the role of these bacteria in improving human intestinal health; however, a number of reports have shown that current total fecal DNA extraction methods and 16S rRNA universal primer sets could affect the species coverage and resolution of these analyses. Here, we improved the extraction method for total DNA from human fecal samples by optimization of the lysis buffer, boiling time (10 min), and bead-beating time (0 min). In addition, we developed a new long-range 16S rRNA universal PCR primer set targeting the V6 to V9 regions with a 580 bp DNA product length. This new 16S rRNA primer set was evaluated by comparison with two previously developed 16S rRNA universal primer sets and showed high species coverage and resolution. The optimized total fecal DNA extraction method and newly designed long-range 16S rRNA universal primer set will be useful for the highly accurate metagenomic analysis of adult and infant intestinal microbiota with minimization of any bias.

Recyclable single-stranded DNA template for synthesis of siRNAs

  • Ali, Mussa M.;Obregon, Demian;Agrawal, Krishna C.;Mansour, Mahmoud;Abdel-Mageed, Asim B.
    • BMB Reports
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    • v.43 no.11
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    • pp.732-737
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    • 2010
  • RNA interference is a post-transcriptional silencing mechanism triggered by the bioavailability and/or exogenous introduction of double-stranded RNA (dsRNA) into cells. Here we describe a novel method for the synthesis of siRNA in a single vessel. The method employs in vitro transcription and a single-stranded DNA (ssDNA) template and design, which incorporates upon self-annealing, two promoters, two templates, and three loop regions. Using this method of synthesis we generated efficacious siRNAs designed to silence both exogenous and endogenous genes in mammalian cells. Due to its unique design the single-stranded template is easily amenable to adaptation for attachment to surface platforms for synthesis of siRNAs. A siRNA synthesis platform was generated using a 3' end-biotinylated ssDNA template tethered to a streptavidin coated surface that generates stable siRNAs under multiple cycles of production. Together these data demonstrate a unique and robust method for scalable siRNA synthesis with potential application in RNAi-based array systems.