• Title/Summary/Keyword: Target DNA

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Quantitative Trait Loci Associated with Functional Stay-Green SNU-SG1 in Rice

  • Yoo, Soo-Cheul;Cho, Sung-Hwan;Zhang, Haitao;Paik, Hyo-Chung;Lee, Chung-Hee;Li, Jinjie;Yoo, Jeong-Hoon;Lee, Byun-Woo;Koh, Hee-Jong;Seo, Hak Soo;Paek, Nam-Chon
    • Molecules and Cells
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    • v.24 no.1
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    • pp.83-94
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    • 2007
  • During monocarpic senescence in higher plants, functional stay-green delays leaf yellowing, maintaining photosynthetic competence, whereas nonfunctional stay-green retains leaf greenness without sustaining photosynthetic activity. Thus, functional stay-green is considered a beneficial trait that can increase grain yield in cereal crops. A stay-green japonica rice 'SNU-SG1' had a good seed-setting rate and grain yield, indicating the presence of a functional stay-green genotype. SNU-SG1 was crossed with two regular cultivars to determine the inheritance mode and identify major QTLs conferring stay-green in SNU-SG1. For QTL analysis, linkage maps with 100 and 116 DNA marker loci were constructed using selective genotyping with $F_2$ and RIL (recombinant inbred line) populations, respectively. Molecular marker-based QTL analyses with both populations revealed that the functional stay-green phenotype of SNU-SG1 is regulated by several major QTLs accounting for a large portion of the genetic variation. Three main-effect QTLs located on chromosomes 7 and 9 were detected in both populations and a number of epistatic-effect QTLs were also found. The amount of variation explained by several digenic interactions was larger than that explained by main-effect QTLs. Two main-effect QTLs on chromosome 9 can be considered the target loci that most influence the functional stay-green in SNU-SG1. The functional stay-green QTLs may help develop low-input high-yielding rice cultivars by QTL-marker-assisted breeding with SNU-SG1.

Tumour Suppressive Effects of WEE1 Gene Silencing in Breast Cancer Cells

  • Ghiasi, Naghmeh;Habibagahi, Mojtaba;Rosli, Rozita;Ghaderi, Abbas;Yusoff, Khatijah;Hosseini, Ahmad;Abdullah, Syahrilnizam;Jaberipour, Mansooreh
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6605-6611
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    • 2013
  • Background: WEE1 is a G2/M checkpoint regulator protein. Various studies have indicated that WEE1 could be a good target for cancer therapy. The main aim of this study was to asssess the tumor suppressive potential of WEE1 silencing in two different breast cancer cell lines, MCF7 which carries the wild-type p53 and MDA-MB468 which contains a mutant type. Materials and Methods: After WEE1 knockdown with specific shRNAs downstream effects on cell viability and cell cycle progression were determined using MTT and flow cytometry analyses, respectively. Real-time PCR and Western blotting were conducted to assess the effect of WEE1 inhibition on the expression of apoptotic (p53) and anti-apoptotic (Bcl2) factors and also a growth marker (VEGF). Results: The results showed that WEE1 inhibition could cause a significant decrease in the viability of both MCF7 and MDA-MB-468 breast cancer cell lines by more than 50%. Interestingly, DNA content assays showed a significant increase in apoptotic cells following WEE1 silencing. WEE1 inhibition also induced upregulation of the apoptotic marker, p53, in breast cancer cells. A significant decrease in the expression of VEGF and Bcl-2 was observed following WEE1 inhibition in both cell lines. Conclusions: In concordance with previous studies, our data showed that WEE1 inhibition could induce G2 arrest abrogation and consequent cell death in breast cancer cells. Moreover, in this study, the observed interactions between the pro- and anti-apoptotic proteins and decrease in the angiogenesis marker expression confirm the susceptibility to apoptosis and validate the tumor suppressive effect of WEE1 inhibition in breast cancer cells. Interestingly, the levels of the sensitivity to WEE1 silencing in breast cancer cells, MCF7 and MDA-MB468, seem to be in concordance with the level of p53 expression.

Expressions of Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptor Gene in the Rat Uterus (흰쥐 자궁에서 Pituitary Adenylate Cyclase-Activating Polypeptide와 수용체 유전자의 발현)

  • 이성호
    • Development and Reproduction
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    • v.2 no.1
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    • pp.21-27
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    • 1998
  • The present study was performed to analyze the gene expressions of pituitary adenylate cyclase-activating polypeptide(PACAP) and its receptor in the rat uterus, a candidate for novel extrahypothalamic source and target. The PACAP cDNA fragments corresponding to the common exon region which is found in both the rat hypothalamus and testis were produced from all tissue samples including the rat uterus by reverse transcriptionpolymerase chain reaction (RT-PCR). No PCR product was amplified from the rat hypothalamic, pituitary, ovarian and uterine samples when the 5' primer corresponding to the testis-specific exon 1 region was used, while the predicted size of product was detected from the testis sample. RT-PCR using the uterine RNA and specific primers for the PACAP receptor yielded products with predicted sizes. Transcripts for the rat uterine PACAP receptor were identified as type I isoforms with hip-hop and hip- or hop-type inserts. After pregnant mare's serum gonadotropin (15 IU) treatment of immature rats (day 25), the level of PACAP mRNA was increased in 24 h and 48 h group, and was declined to the lowest in 72 h group. The present study shows the presence of transcripts for PACAP and its receptor isoform in the rat uterus. These finding ssuggest that the uterine PACAP ight act as a novel autocrine and/or paracrine factor via its specific receptors on the reglulation of rat uterine function and physiology during the reproductive cycle.

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Molecular Analysis of Pathogenic Molds Isolated from Clinical Specimen (임상검체에서 분리된 병원성 사상균의 분자생물학적 분석)

  • Lee, Jang Ho;Kwon, Kye Chul;Koo, Sun Hoe
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.229-236
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    • 2020
  • Sixty-five molds isolated from clinical specimens were included in this study. All the isolates were molds that could be identified morphologically, strains that are difficult to identify because of morphological similarities, and strains that require species-level identification. PCR and direct sequencing were performed to target the internal transcribed spacer (ITS) region, the D1/D2 region, and the β-tubulin gene. Comparative sequence analysis using the GenBank database was performed using the basic local alignment search tool (BLAST) algorithm. The fungi identified morphologically to the genus level were 67%. Sequencing analysis was performed on 62 genera and species level of the 65 strains. Discrepancies were 14 (21.5%) of the 65 strains between the results of phenotypic and molecular identification. B. dermatitidis, T. marneffei, and G. argillacea were identified for the first time in Korea using the DNA sequencing method. Morphological identification is a very useful method in terms of the reporting time and costs in cases of frequently isolated and rapid growth, such as Aspergillus. When molecular methods are employed, the cost and clinical significance should be considered. On the other hand, the molecular identification of molds can provide fast and accurate results.

Validation of a HPLC MS/MS Method for Determination of Doxorubicin in Mouse Serum and its Small Tissues (마우스 혈장과 조직에서의 doxorubicin 측정 HPLC-MS/MS 방법)

  • Park, Jung-Sun;Kim, Hye-Kyung;Lee, Hye-Won;Lee, Mi-Hyun;Kim, Hyun-Gi;Chae, Soo-Wan;Chae, Han-Jung
    • Korean Journal of Clinical Pharmacy
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    • v.16 no.1
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    • pp.23-27
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    • 2006
  • Doxorubicin (DXR) is a type of anti-cancer drug called an 'anthracycline glycoside', It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at $350{\mu}L/min$. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.

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Effect of Feed Types on Ochratoxin A Disappearance in Goat Rumen Fluid

  • Upadhaya, Santi Devi;Yang, Liu;Seo, Ja-Kyeom;Kim, Myung-Hoo;Lee, Chang-Kyu;Lee, Chan-Ho;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.2
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    • pp.198-205
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    • 2011
  • This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.

Polymorphism of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Native Sarabi Cows

  • Pashmi, M.;Ghorashi, S.A.;Salehi, A.R.;Moini, M.;Javanmard, A.;Qanbari, S.;Yadranji-Aghdam, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.775-778
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    • 2006
  • Sarabi cows (n = 136) from the Sarabi Breeding Station were genotyped at bovine lymphocyte antigen (BoLA)-DRB3.2 locus by a genotyping system that used the polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from whole blood samples. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of target gene. Nested-PCR products were digested with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamide gel electrophoresis. Twenty-six BoLA-DRB3.2 alleles were identified with frequencies ranging from 0.4 to 15.1%. Six new allele types observed in this study have not been reported previously. Identified alleles include: BoLA-DRB3.$2^*1$, $^*2$, $^*4$, $^*6$, $^*8$, $^*12$, $^*13$, $^*14$, $^*15$, $^*16$, $^*17$, $^*23$, $^*24$, $^*25$, $^*28$, $^*32$, $^*34$, $^*35$, $^*36$, $^*37$, $^*42$, $^*46$, $^*51$, $^*kba$, $^*laa$ and $^*vaa$. Their frequencies were found to be 0.4, 0.4, 0.7, 11.4, 1.1, 1.8, 2.9, 2.2, 4.4, 9.6, 1.1, 13.6, 0.4, 0.4, 1.1, 0.7, 0.4, 6.2, 2.2, 3.7, 1.1, 7.7, 1.5, 15.1, 2.6 and 7.3% respectively. The six most frequent alleles (DRB3.2 $^*6$, $^*16$, $^*23$, $^*46$, $^*kba$ and $^*vaa$) accounted for 64.7% of the alleles in the population of this herd. Numerous studies on this locus, covering different breeds, has revealed the existence of various alleles in this locus, and new investigations have introduced novel alleles. With respect to the high number of the observed alleles in this survey and the novelty of some alleles with no previous record of reporting, it is plausible to conclude that the BoLA-DRB3.2 locus is highly polymorphic in Iranian native Sarabi cows.

Transcription Factor for Gene Function Analysis in Maize (옥수수 유전자 기능 분석을 위한 전사인자의 이해)

  • Moon, Jun-Cheol;Kim, Jae Yoon;Baek, Seong-Bum;Kwon, Young-Up;Song, Kitae;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.263-281
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    • 2014
  • Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3/VP1, AP2/EREBP, ARF, ARID, AS2, AUX/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2-YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.

Effect of Gamisamgibopae-tang on the Growth and Apoptosis of A539 and NCI-H460 Human Lung Cancer Cells (A539 및 NCI-H460 인체 폐암세포의 증식 및 apoptosis 유도에 미치는 가미삼기보폐탕의 영향)

  • Kim, Jin-Young;Kim, Hyun-Joong;Jung, Kwang-Sik;Park, Cheol;Choi, Yung-Hyun;Kam, Cheol-Woo;Park, Dong-Il
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.130-148
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    • 2008
  • Objective : This study was designed to investigate the effect of the water extract of Gamisamgibopae-tang(GMSGBPT), an oriental herbal formulation, on the growth of NCI-H460 and A549 human non-small-cell lung cancer cell lines. Methods : Cytotoxicity and cell morphology were evaluated by MTT assay and inverted microscope, respectively. Apoptosis was detected using agarose gel electrophoresis and flow cytometer. The expression levels of mRNAs and proteins of target genes were determined by RT-PCR and western blot analyses, respectively Result and Conclusion : We found that exposure of A549 cells to GMSGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, but GMSGBPTdid not affect the growth of NCI-H460 cells. The anti-proliferative effect of GMSGBPT treatment in A549 cells was associated with morphological changes, formation of apoptotic bodies and DNA fragmentation, and flow cytometry analysis confirmed that GMSGBPT treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptotic cell death by GMSGBPT were connected with a up-regulation of cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) mRNA and protein in a tumor suppressor p53-independent fashion. However GMSGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1), inducible nitric oxide synthase (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 orNCI-H460 cells. Taken together, these findings partially provide novel insights into the possible molecular mechanism of the anti-cancer activity of GMSGBPT.

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Evaluation of Sleep Comfort for Indoor Thermal Environment based on the Physiological Signal Analysis (생리신호 분석을 이용한 실내온열환경에서의 수면 쾌적성 평가)

  • 이낙범;임재중;금종수;이구형;최호선
    • Science of Emotion and Sensibility
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    • v.3 no.2
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    • pp.75-84
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    • 2000
  • 문명의 발달과 함께 수면부족으로 인한 여러 가지 스트레스와 질환이 증가하게 되어 최근 수면연구에 대한 관심이 증가하고 있다. 본 연구는 다양한 온열환경 조건에서의 쾌적한 수면을 위한 온열환경을 제시하기 위해, 5명의 여성 피험자를 대상으로 22$^{\circ}C$, 26$^{\circ}C$, 3$0^{\circ}C$의 일정온도 조건과 $25^{\circ}C$에서 1시간 후와 2시간 후에 각각 1, 2$^{\circ}C$를 상승시켜주는 변동온도 조건하에서 수면생리신호를 측정하였다. 그리고, 수면단계 평가를 이용하여 총 수면시간, 깊은 수면의 비율, 그리고 최초 수면시작 시간에서 최초의 서파 수면이 나타나기까지의 지연시간 등의 수면효율을 평가하였다. 그 결과, 일정온도 조건에서는 26$^{\circ}C$에서 총 수면시간(466.7$\pm$10.25분)과 깊은 수면의 비율(33.1$\pm$4.95%)은 타 조건에 비해 높게 나타났고, 최초 서파수면까지의 지연시간(9.8$\pm$3.33분)은 타 조건에 비해 낮게 나타나 쾌적한 수면을 위한 가장 적절한 온열조건임을 관찰할 수 있었다. 그리고 변동온도 조건에서는 4가지 온열조건간에는 큰 차이가 나타나지 않았지만, 모든 조건에서 일정온도 조건보다는 좋은 결과를 나타내었다. 또한 수면 중 신체 움직임과 설문 분석에서도 동일한 결과를 보였다. 본 연구를 통해, 수면생리신호를 이용한 수면 쾌적성 분석은 수면의 질적인 상태를 관찰하는데 매우 적합한 파라메터를 도출할 수 있으며, 여러 가지 수면환경 조건을 평가하는데 매우 유용한 지표가 될 수 있음을 보였다.e results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG./TEX>이었으며, 이는 화성 기원을 지시한다.sucrose를 이용한 2단계 희석이 수정란의 생존성을 향상시키는 것으로 나타났다. 또한 발육 단계별 생존성에 있어서는 발육이 진전된 확장배 반포 시에 동결하는 것이 배반포기에 동결하는 것 보다 유리한 것으로 나타났다.ody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다..ments of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to

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