• 제목/요약/키워드: PCR-PP

검색결과 34건 처리시간 0.03초

수분조절제로서 팽화왕겨를 이용한 계분 발효 특성 및 시비 효과 (Composting and Fertilizing Characteristics of Poultry Manure Mixture with Compressed Expansion Rice Hull as Bulking Agent)

  • 김영선;조성현;이훈수;이긍주
    • 유기물자원화
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    • 제28권4호
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    • pp.5-13
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    • 2020
  • 본 연구는 수분조절제로서 팽화왕겨(compressed expansion rice hull; CERH) 처리에 따른 계분의 발효 특성 및 발효된 가공계분의 시비에 따른 상추와 청경채의 생육에 미치는 영향을 조사하기 위해 수행되었다. 실험에 사용된 처리구는 톱밥 처리구(계분 90%+톱밥10%; SP), 피트모스 처리구(계분 90%+피트모스10%; PP), 팽화왕겨 1 처리구[계분 90%+CERH1 (��1.3 mm) 10%; PCR1] 및 팽화왕겨 2 처리구[계분 90%+CERH2 (��3.0 mm); PCR2]로 설정하였다. 31일간 진행된 계분 발효 과정에서 발효 더미의 온도, 수분 함량, pH 및 총탄소 함량은 수분조절제의 종류에 따른 차이를 나타내지 않았다. 시험 종료 시 PP 처리구와 PCR1 처리구의 질소 함량은 SP 처리구나 PCR2 처리구보다 높았다. 발효 후 건조된 가공계분의 질소, 인산, 칼리 함량은 각각 2.76~3.65%, 1.69~2.05%, 2.53~2.90%로 나타났다. 수분조절제별 가공계분의 작물 재배 시험에서 SP 처리구와 비교할 때, 상추는 PP와 PCR1 처리구에서 10% 이상 건물중이 증가하였으나, 청경채는 PP 처리구에서만 증가되었다. 상기 결과들을 종합할 때, 1.3 mm 크기의 팽화왕겨는 계분 발효 시 수분조절제로서 피트모스나 톱밥을 대신하여 사용할 수 있었다.

위암에서 정량적 역전사 중합효소연쇄반응을 이용한 다중 표지자 분석 (Multiple Genetic Marker Analysis with Using Quantitative RT-PCR in Gastric Cancer)

  • 유문원;이혁준;최수민;유지은;허근;김영국;양한광
    • Journal of Gastric Cancer
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    • 제7권2호
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    • pp.59-66
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    • 2007
  • 목적: 위암 세포주 및 조직에서 다중 표지자 mRNA 발현을 정량적 RT-PCR 검사를 통해 확인함으로써 이들 표지자를 이용하여 위암의 복강내 미세전이 진단이 가능한가를 평가하고자 본 연구를 시행하였다. 대상 및 방법: 12개의 인체 위암 세포주와 10개의 위암 조직을 대상으로 Carcinoembryonic antigen (CEA), Cytokeratin 20 (CK20), Dopa decarboxylase (DDC), L-3-phosphoserine phosphatase (L3PP)의 네 가지 mRNA를 이용한 정량적 RT-PCR 다중 표지자 분석을 시행하였다. 결과: 12개의 인체 위암 세포주 중 CEA는 4개(33%), CK-20는 1개(8%), DDC는 6개(50%), L3PP는 12개 세포주 모두(100%)에서 과발현되었다. 10개의 위암 조직 중 CEA는 9개, CK20은 3개, DDC는 9개, L3PP는 10개 조직 모두에서 과발현되었다. L3PP는 모든 위암 세포주와 조직에서 과발현을 나타내었으나 과발현 정도는 비교적 낮게 측정된 반면, CEA와 DDC는 일부 위암 세포주 및 조직에서만 과발현을 나타내었지만 파발현 시 충분한 발현도를 나타내었다. 결론: 위암 환자에서 하나 이상의 암 특이적 유전자를 이용한 다중 표지자 분석은 단일 표지자 분석이 가지는 단점을 보완할 수 있을 것으로 예상되며, CEA, DDC, 및 L3PP의 세 가지 mRNA가 후보 유전자로 사용될 수 있을 것으로 생각한다.

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Development of Multiplex PCR for Simultaneous Detection of Citrus Viruses and the Incidence of Citrus Viral Diseases in Late-Maturity Citrus Trees in Jeju Island

  • Hyun, Jae Wook;Hwang, Rok Yeon;Jung, Kyung Eun
    • The Plant Pathology Journal
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    • 제33권3호
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    • pp.307-317
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    • 2017
  • Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RTPCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5-96.2% and 97.1-97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.

류마티스 관절염에서 HO-1에 의한 VEGF 발현 유도에 대한 연구 (Effects of Heme Oxygenase-1 on VEGF Expression in Rheumatoid Arthritis)

  • 이승훈;권상철;변승재;장성조
    • 동의생리병리학회지
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    • 제22권4호
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    • pp.871-877
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    • 2008
  • Heme oxygenase-1 (HO-1), an inducible heme-degrading enzyme, is expressed by macrophages and endothelial cells in response to various stresses and mediators of inflammation. HO-1 has been recently implicated in regulation of angiogenesis via expression of VEGF. The purpose of this study was to determine the effects of HO-1 modulation on the collagen-induced arthritis (CIA) model and on angiogenesis via up- regulation of VEGF expression in human synovial fibroblast. DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 1 to day 35 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, histopathologic changes were examined on the joints. VEGF expression in paws were measured by immunohistochemical stain. mRNA expression of HO-1 and VEGF stimulated with various concentration of $TNF-{\alpha}$, CoPP accessed on human synovial fibroblast by RT-PCR. Effects of pretreatment with SnPP on mRNA expression of HO-1 and VEGF in the presence of CoPP and $TNF-{\alpha}$ in synovial fibroblast was accessed by Real-time RT-PCR. Administration of cobalt protoporphyrin IX significantly induced the inflammatory response, with increased arthritis index and expression of VEGF in the paws of the arthritis models. Treatment with SnPP significantly reduced the severity of CIA through inhibition of joint inflammation and cartilage destruction. The expression of VEGF were also significantly reduced by SnPP treatment in the paw. CoPPIX as inducer of HO-1, increased HO-1 and VEGF expression dose dependently in synovial fibroblast. In contrast, inhibition of HO-1 activity by SnPPIX abrogated CoPPIX-induced HO-1 and VEGF production in synovial fibroblast. Stimulation with $TNF-{\alpha}$ increased HO-1 and VEGF expression itself and showed additive effect on HO-1 and VEGF expression when it treated with CoPP. When SnPP was treated with CoPP and $TNF-{\alpha}$, it abrogated the CoPP induced HO-1 and VEGF expression and also abrogated $TNF-{\alpha}$ induced HO-1 and VEGF expression in synovial fibroblast. The effects of HO-1 induction in rheumatoid arthritis results in aggravation of arthritis via up-regulation of VEGF. I concluded that inhibition of the expression or activity of HO-1 could be a therapeutic target of rheumatoid arthritis.

Paralytic Peptide Binding Protein (PP-BP) Gene Expression During Egg Diapause and Its Multi-Gene Organization in the Silkworm Bombyx mori

  • Sirigineedi, Sasibhushan;Murthy, Geetha N.;Rao, Guruprasada;Ponnuvel, Kangayam M.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제26권1호
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    • pp.31-40
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    • 2013
  • Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • 제34권8호
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Genetic Variability among Different Populations of Root Knot Nematodes Based on Their Encumbrance Response to Pasteuria Isolates Using PCR-RFLP

  • Kamran, Muhammad;Javed, Nazir;Ullah, Ihsan;Nazir, Shahid;Jamil, Shakra;Iqbal, Muhammad Zafar;Abbas, Huma;Khan, Sajid Aleem;Haq, Muhammad Ehetisham ul
    • The Plant Pathology Journal
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    • 제35권1호
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    • pp.51-62
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    • 2019
  • A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures ($25{\pm}2^{\circ}C$ and $30{\pm}2^{\circ}C$) and concentrations ($10^4$ and $10^5$ spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.

Evaluation of JC and Cytomegalo Viruses in Glioblastoma Tissue

  • Afshar, Reza Malekpour;Mollaei, Hamid Reza;Zandi, Bahare;Iranpour, Maryam
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권11호
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    • pp.4907-4911
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    • 2016
  • Glioblastoma multiforme (GBM) is the most aggressive of the gliomas, a collection of tumors arising from glia in the central nervous system. Possible associations between the human cytomegalovirus (HCMV) and the JC virus with GBM are now attracting interest. Our present aim was to investigate the prevalence of the two viruses in Iranian patients from Kerman's cities in the south of Iran. In addition, the expression rates of pp65, large T antigen and p53 proteins were assessed and their relation with GBM evaluated using reverse transcription real time PCR (rReal Time PCR). A total of 199 patients with GBM cancer were enrolled, with $mean{\pm}SD$ ages of $50.0{\pm}19.5$ and $50.7{\pm}19.6$ years for males and females, respectively. The P53 rate was dramatically low suggesting an aetiological role,. Large T antigen expression was found in JC positive samples, while the PP65 antigen was observed in patients positive for CMV and JC. HCMV products and JC virus with oncogenic potential may induce the development of various tumors including glioblastomas. The JC virus produces an early gene product, T-antigen, which has the ability to associate with and functionally inactivate well-studied tumor suppressor proteins including p53 and pRB.

마우스 내장 림프조직에서 우세하게 발현되는 IgA Isotype Switching 관련 전사체의 분석 (Preferential Expression of IgA Isotype Switching-associated Transcripts in Mouse Intestinal Lymphoid Tissues)

  • 채병철;전성기;서구영;김현아;김평현
    • IMMUNE NETWORK
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    • 제5권4호
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    • pp.215-220
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    • 2005
  • Background: Transforming growth factor-${\beta}$ (TGF-${\beta}1$) directs class switch recombination (CSR) to IgA isotype, which is a predominant antibody in mucosal surfaces. Although IgA is preferentially committed in mucosal lymphoid tissues, it is not definitely established whether hallmarks of IgA CSR such as IgA germ-line transcripts (GLT ${\alpha}$), post-switch transcripts (PST ${\alpha}$) and circle transcripts (CT ${\alpha}$) are readily expressed in such tissues. Therefore, we compared the expression of these transcripts among mouse Peyer's patches (PP), mesenteric lymph nodes (MLN), and spleen. Methods: Levels of GLTs, PSTs and CTs were measured by RT-PCR in isolated PPs, MLNs and spleen cells. Results: GLT ${\alpha}$ and PST ${\alpha}$ were well expressed in PP and MLN cells but in spleen cells. Similar patterns were observed in the expression of GL ${\gamma}$2b and PST ${\gamma}$2b. On the other hand, these transcripts were only inducible in spleen cells upon stimulated with LPS and TGF-${\beta}1$. In addition, CT${\alpha}$ and CT${\gamma}$2b were detected in PP cells. Conclusion: PP B cells readily express IgA GLT, PST, and CT. Overall expression patterns of these transcripts were similar in MLN cells. Thus, these results suggest that microenvironment of PP and MLN influences spontaneous IgA CSR, which lacks in systemic lymphoid tissues such as spleen.

재활용 플라스틱 펠렛과 벨벳콩(Mucuna pruriens) 추출물의 항산화 효능 상관관계 검증 (Verification of Recycled Plastic Pellets and Mucuna pruriens Extracts Correlation of Antioxidant Efficacy)

  • 한동근;배민준;갈격;김현정;김세기;안봉전
    • 대한화장품학회지
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    • 제49권1호
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    • pp.29-38
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    • 2023
  • 본 연구에서는 재활용 플라스틱 펠렛과 벨벳콩(Mucuna pruriens) 추출물의 항산화 효능 안정도에 대한 상관관계를 검증하였다. 재활용 플라스틱 펠렛을 벨벳콩 추출물에 침지 시킨 다음 다양한 온도 조건에서 30일간 보관한 후 DPPH radical 및 ABTS+ radical 소거능, SOD 유사 활성, 총 페놀 및 플라보노이드 함량 분석을 실시하였다. 기존 효능에 대비하여 DPPH radical 소거능은 ECOTRIA, PCR-PP 펠렛에서 30% 미만의 활성변화를 나타내었으며, Grico 펠렛에서 46.06%의 가장 큰 활성 감소가 확인되었다. ABTS+ radical 소거능 측정결과, 모든 펠렛에서 5% 미만의 효능 변화가 확인되었다. SOD 유사 활성을 측정한 결과, 모두 30% 이상의 활성 감소를 나타내었으며, 총 페놀 및 플라보노이드 함량 측정 결과, 모든 펠렛에서 기존 함량 대비 감소하는 경향을 나타내었다. 이를 통해 재활용 플라스틱 펠렛에 의한 벨벳콩 추출물의 생리활성 감소 및 변화를 확인하였으며, 본 연구 결과를 토대로 재활용 플라스틱 펠렛은 벨벳콩 추출물이 지닌 기존 효능 및 활성의 안정도에 영향을 미치는 것으로 판단된다.