• 제목/요약/키워드: RNA I

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흰쥐 성주기간동안 Prolactin mRNA의 변화:Naloxone (Alterations in Prolactin Messenger Ribonucleic Acid Level During the Rat Estrous Cycle: Effect of Naloxone)

  • 안혜영;유선경;조병남;김경진;유경자;조완규
    • 한국동물학회지
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    • 제33권2호
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    • pp.183-190
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    • 1990
  • 본 연구는 prolactin(PRL)유전자 발현, 분비의 생리적 변화와 성주기 특정 시기의 PRL mRNA수준 및 분비에 미치는 내인성 오피오이드의 영향을 조사하였다. 최소한 두번의 연속적인 성주기를 거친 성숙한 흰쥐에서 성주기의 각 시기(10:00시)에, proestrus시기에는 10:00-20:00 시동안에는 2시간 간격으로 도살하였고, naloxone (2mg/kg b.w.)은 도살 30분전에 피하주사 하였다. PRL mRNA의 수준의 흰쥐의 PRL cDNA를 probe로 하여 RNA-blot hybridization방법에 의해서, 혈중 PRL농도 변화는 방사면역측정법에 의해 측정하였다. 뇌하수체 PRL mRNA의 수준과 혈중 PRL수준은 diestrus I, II and proestrus그리고 estrus시기의 10:00시에는 급격한 변화를 보이이 않았다. 이때 naloxone처리는 영향을 미치지 못했다. proestrus시기를 세분화하여 조사한 결과 PRL mRNA의 수준은 정오에 최고 수준에 도달하였고, 오후 6:00까지 점차적으로 감소하였다. 그후 8:00시에 다시 증가하였다. estrus동안 naloxone은 혈중의 PRL수준을 명백히 억제하였으나 PRL mRNA수준에는 영향이 없었다. proestrus시기 동안 혈중 PRL변화와 뇌하수체 PRL mRNA변화는 서로 상이하게 조절되며,PRL mRNA수준이 흰쥐 성주기 동안 변화하고 있는 사실에서 PRL 유전자 발현이 생리적으로 조절되고 있음을 시사한다.

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The First Identified Citrus tristeza virus Isolate of Turkey Contains a Mixture of Mild and Severe Strains

  • Cevik, Bayram;Yardimci, Nejla;Korkmaz, Sava
    • The Plant Pathology Journal
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    • 제29권1호
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    • pp.31-41
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    • 2013
  • The presence of Citrus tristeza virus (CTV) has previously been reported in citrus growing regions of Turkey. All serologically and biologically characterized isolates including I$\breve{g}$d${\i}$r, which was the first identified CTV isolates from Turkey, were considered mild isolates. In this study, molecular characteristics of the I d r isolate were determined by different methods. Analysis of the I$\breve{g}$d${\i}$r isolate by western blot and BD-RT-PCR assays showed the presence of MCA13 epitope, predominantly found in severe isolates, in the I$\breve{g}$d${\i}$r isolate revealing that it contains a severe component. For further characterization, the coat protein (CP) and the RNA-depen-dent RNA polymerase (RdRp) genes representing the 3' and 5' half of CTV genome, respectively, were amplified from dsRNA by RT-PCR. Both genes were cloned separately and two clones for each gene were sequenced. Comparisons of nucleotide and deduced amino acid sequences showed that while two CP gene sequences were identical, two RdRp clones showed only 90% and 91% sequence identity in their nucleotide and amino acid sequences, respectively, suggesting a mixed infection with different strains. Phylogenetic analyses of the CP and RdRp genes of I$\breve{g}$d${\i}$r isolate with previously characterized CTV isolates from different citrus growing regions showed that the CP gene was clustered with NZRB-TH30, a resistance breaking isolate from New Zealand, clearly showing the presence of severe component. Furthermore, two different clones of the RdRp gene were clustered separately with different CTV isolates with a diverse biological activity. While the RdRp-1 was clustered with T30 and T385, two well-characterized mild isolates from Florida and Spain, respectively, the RdRp-2 was most closely related to NZRB-G90 and NZRB-TH30, two well-characterized resistance breaking and stem pitting (SP) isolates from New Zealand confirming the mixed infection. These results clearly demonstrated that the I$\breve{g}$d${\i}$r isolate, which was previously described as biologically a mild isolate, actually contains a mixture of mild and severe strains.

Hepatic Vascular Stress Gene Expression in the Liver Response to Trauma

  • Lee, Sun-Mee
    • Biomolecules & Therapeutics
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    • 제12권2호
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    • pp.62-67
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    • 2004
  • Trauma remains one of the important sources leading to systemic inflammatory response anti sub-sequent multiple organ failure. Although hepatic microvascular dysfunction occurs during trauma, the mechanism responsible remains unclear. The aim of this study was to investigate the effect of trauma on hepatic vascular stress gene expression. Femur fracture (EFx) was induced by torsion to the femur at midshaft. Liver samples were taken for RT-PCR analysis of mRNA for gtenes of interest: endothelin-1 (ET-1), its receptors $ET_A$ and $ET_B$, nitric oxide synthases (iNOS and eNOS), cyclooxygenase-2 (COX-2), heme oxygenase-1 (HO-1), and tumor necrosis tactor-${\alpha}$ (TNF-${\alpha}$). The expression of ET-1 mRNA was significantly increased by FFx. Expression of mRNA in FFx group showed no change in $ET_A$, $ET_B$, iNOS and HO-1 and showed a slight increase of 2.2-fold and 2.7-fold for eNOS tll1d COX-2, respectively. The level of TNF-${\alpha}$ mRNA significantly increased in FFx group. In conclusion, mild trauma alone causes little change in expression of vasoactive mediators.

Electrical Stimulation Promotes Healing Accompanied by NOR in Keratinocytes and IGF-1 mRNA Expression in Skin Wound of Rat

  • Lee, Jae-Hyoung;Lee, Jong-Sook;Jeong, Myung-A.;JeKal, Seung-Joo;Kil, Eyn-Young;Park, Seung-Teack;Park, Chan-Eui
    • 대한의생명과학회지
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    • 제13권1호
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    • pp.25-32
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    • 2007
  • The purpose of this study was to investigate the effect of the high voltage pulsed Current (HVPC) stimulation on the healing rate and the proliferative activity of keratinocytes and IGF-I mRNA expression of an incisional wound in rat skin. Twenty male Sprague-Dawley rats ($265{\sim}290g$) were randomly divided into HVPC (n=10) and control group (n=10). Rats received 10 mm length of full-thickness incision wound on the back under the anesthesia. The HVPC group received electrical stimulation with a Current intensity of 50 V at 100 pps for a duration of 30 minutes, while the control group was given the same treatment without electricity for a week. Polarity was negative in first three days and positive thereafter. The wound length was measured and evaluated as percentage. The mean number of nucleolar organizer regions (NORs) per nucleus and level of IGF-I mRNA expression were calculated. The mean percent of wound closure were $51.17{\pm}17.76%$ and $80.71{\pm}11.91%$, respectively, in the sham treated control and HVPC stimulated groups (t=-4.308, P<0.001). The mean NOR number per nucleus of the keratinocytes in the control and HVPC group were $1.85{\pm}0.20$ and $2.70{\pm}0.23$, respectively (t=8.638, P<0.001). The IGF-I mRNA level were $0.76{\pm}0.44$ and $1.32{\pm}0.41$, respectively, in the control and HVPC stimulated wounds (t=2.906, P<0.01). There was a positive correlation between the mean NOR number per nucleus and IGF-l mRNA level with a Pearson product moment correlation coefficient of 0.72 (P<0.05). These findings suggest that the HVPC may activate the rRNA of the basal keratinocytes and upregulate the IGF-I mRNA levels by alteration of the electrical environment, and it may increase proliferative activity of the keratinocytes in the skin wound of the rat.

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미토콘드리아 12S rRNA 유전자의 종 특이적 PCR-RFLP Fingerprint를 이용한 식육 원료의 판별 (Identification of Meat Species Using Species-Specific PCR-RFLP Fingerprint of Mitochondrial 12S rRNA Gene)

  • 박종근;신기현;신성철;정구용;정의룡
    • 한국축산식품학회지
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    • 제27권2호
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    • pp.209-215
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    • 2007
  • 본 연구는 mt DNA 12S rRNA 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 9종류 축종(소, 돼지, 양, 염소, 말, 사슴, 닭, 오리 및 칠면조)의 육류로부터 12S rRNA유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 공시축의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(약 455 bp)을 Tsp5091와 MboI 제한효소로 각각 절단한 결과 Tsp5091 제한효소는 포유류 6종간에서 그리고 MboI 제한효소는 가금류 3종간에서 명확한 차이를 보이는 종 특이적인 PCR-RFLP profile을 검출하였다. 따라서 본 연구에서 개발한 12S rRNA 유전자의 종 특이적 DNA 분자표지는 각종 원료육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 동물 종 감별 DNA marker로 이용될 수 있을 것이다.

As계의 오이 모자이크 바이러스 RNA4의 염기서열 결정 (Determination of Nucleotide Sequences of cDNA from Cucumber Mosaic Virus-As RNA4)

  • 김상현;박원목;이세영;박영인
    • 한국식물병리학회지
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    • 제12권2호
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    • pp.176-181
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    • 1996
  • Aster yomena로부터 분리한 오이 모자이크 바이러스(cucumber mosaic virus) (CMV-As)의 RNA4로부터 완전한 길이의 cDNA를 합성하고 그 전체적인 염기서열(1,043 nt`s)을 결정하였다. CMV-As RNA4는 73개의 염기로 구성된 5`말단의 leader 부위, 657개의 염기로 구성된 외피단백질(coat protein) 유전자 부위 및 312개의 염기로 구성된 3` 말단의 비번역 부위로 구성되어 있음을 확인하였다. 외피단백질 유전자 부위의 염기서열을 다른 계통의 CMV와 비교해 볼 때 그 염기서열이 보전적으로 존재하고 있으나 그 외의 부분은 다양함을 확인하였다. 특히 3` 말단부위의 61개의 염기로 구성된 부위(959-1019)는 다른 계통의 CMV에서는 상당히 유사하지만 CMV-As도 다른 CMV처럼 tRNA와 유사한 구조를 역시 형성함을 확인하였다. CMV-As의 RNA4 염기서열을 다른 계통의 CMV와 비교할 때 CMV-I17F와 가장 유사하였으며(91.9%) S형의 CMV-M과는 가장 낮은 동일성을 보였다(71.1%). 외와 같은 염기성열의 비교 결과와 EcoRI 제한효소 인식부위의 존재로 미루어 CMV-As는 WT형으로 분류된다.

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An RNA Mapping Strategy to Identify Ribozyme-Accessible Sites on the Catalytic Subunit of Mouse Telomerase

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • 제5권1호
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    • pp.32-35
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    • 2007
  • Telomerase reverse transcriptase (TERT) is an enzymatic ribonucleoprotein that prolongs the replicative life span of cells by maintaining protective structures at the ends of eukaryotic chromosomes. Telomerase activity is highly up-regulated in 85-90% of human cancers, and is predominately regulated by hTERT expression. In contrast, most normal somatic tissues in humans express low or undetectable levels of telomerase activity. This expression profile identifies TERT as a potential anticancer target. By using an RNA mapping strategy based on a trans-splicing ribozyme library, we identified the regions of mouse TERT (mTERT) RNA that were accessible to ribozymes. We found that particularly accessible sites were present downstream of the AUG start codon. This mTERTspecific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors.

Dimensionality Reduction of RNA-Seq Data

  • Al-Turaiki, Isra
    • International Journal of Computer Science & Network Security
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    • 제21권3호
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    • pp.31-36
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    • 2021
  • RNA sequencing (RNA-Seq) is a technology that facilitates transcriptome analysis using next-generation sequencing (NSG) tools. Information on the quantity and sequences of RNA is vital to relate our genomes to functional protein expression. RNA-Seq data are characterized as being high-dimensional in that the number of variables (i.e., transcripts) far exceeds the number of observations (e.g., experiments). Given the wide range of dimensionality reduction techniques, it is not clear which is best for RNA-Seq data analysis. In this paper, we study the effect of three dimensionality reduction techniques to improve the classification of the RNA-Seq dataset. In particular, we use PCA, SVD, and SOM to obtain a reduced feature space. We built nine classification models for a cancer dataset and compared their performance. Our experimental results indicate that better classification performance is obtained with PCA and SOM. Overall, the combinations PCA+KNN, SOM+RF, and SOM+KNN produce preferred results.

Investigation of functional roles of transcription termination factor-1 (TTF-I) in HIV-1 replication

  • Park, Seong-Hyun;Yu, Kyung-Lee;Jung, Yu-Mi;Lee, Seong-Deok;Kim, Min-Jeong;You, Ji-Chang
    • BMB Reports
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    • 제51권7호
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    • pp.338-343
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    • 2018
  • Transcription termination factor-1 (TTF-I) is an RNA polymerase 1-mediated transcription terminator and consisting of a C-terminal DNA-binding domain, central domain, and N-terminal regulatory domain. This protein binds to a so-called 'Sal box' composed of an 11-base pair motif. The interaction of TTF-I with the 'Sal box' is important for many cellular events, including efficient termination of RNA polymerase-1 activity involved in pre-rRNA synthesis and formation of a chromatin loop. To further understand the role of TTF-I in human immunodeficiency virus (HIV)-I virus production, we generated various TTF-I mutant forms. Through a series of studies of the over-expression of TTF-I and its derivatives along with co-transfection with either proviral DNA or HIV-I long terminal repeat (LTR)-driven reporter vectors, we determined that wild-type TTF-I downregulates HIV-I LTR activity and virus production, while the TTF-I Myb-like domain alone upregulated virus production, suggesting that wild-type TTF-I inhibits virus production and trans-activation of the LTR sequence; the Myb-like domain of TTF-I increased virus production and trans-activated LTR activity.