• 제목/요약/키워드: universal primers

검색결과 105건 처리시간 0.025초

다래나무속 식물의 분류 및 계통 특이밴드 탐색을 위한 범용 프라이머 개발 (Development of Universal Primers for Phylogenetic Analysis and Species-specific Band Identification in the Genus Actinidia)

  • 김성철;장기창;송은영;김공호;정용환;김미선;오순자;고석찬
    • 한국자원식물학회지
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    • 제17권2호
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    • pp.107-115
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    • 2004
  • 참다래 육종을 위한 종 분류와 분자 표지인자로서 유용하게 이용될 수 있는 primer를 개발하기 위하여 참다래 genome 특이 반복 염기서열로부터 19∼20base 크기로 18개의 primer를 제작하여 kiwifruit target primer(KT primer)라 명명하였으며, 동아시아 지역에서 수집된 7종 22계통의 다래나무 속 식물을 이용하여 활용 가능성 을 조사하였다. 유연관계 분석을 위하여 7개의 primer가 선발되었으며, 이를 이용한 RAPD 결과 크게 2개의 군으로 나뉘어 졌다. 제 1 군(A. arguta, A. melanandra, A. kolumikta와 A. marcrosperma)은 주로 과실에 전혀 털이 없으며 잎에는 털이 전혀 없거나 어렸을 때 극소량의 연모가 있다가 없어지는 그룹으로서 Leiocarpae 절에 속하였다. 제 2 군(A. chinensis, A. deliciosa 및 A. eriantha)은 어린 과실에서는 털이 많았다가 성숙하면서 털이 없어지는 계통 및 잎과 줄기에 털이 아주 많거나 조밀한 솜털이 있는 그룹으로서 Stellatae절에 속하였다. 제 2 군은 Stellatae 절에서도 Pefectae 아절에 속하는 것으로 A. chinensis, A. deliciosa 및 A. eriantha가 포함되었으며, 다시 A. chinensis와 A. deliciosa를 포함하는 그룹과 A. eriantha 등 2개의 그룹으로 나뉘어졌다. 같은 부모로부터 유래된 것으로 알려진 A. chinensis와 A. deliciosa는 80%의 유사도에서 두 개의 그룹으로 나뉘어졌다. 또한 PCR 결과 A. deliciosa 종 및 헤이워드와 토무리 계통 특이 밴드가 KT12F와 KT6F에서 나타났으며, 유전양상 분석에서 KT7F와 KT12F가 유용하였다. 본 연구 결과 KT primer는 참다래의 유전양상 분석과 특이한 유전양상을 나타내는 개체선발 및 도태에 유용하게 이용될 수 있고, 또한 참다래 육종 효율향상에 많은 도움을 줄 수 있다고 판단되었다.

Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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니켈-크롬 합금에 대한 다용도 접착 시스템의 전단결합강도 (Shear bond strength of Universal bonding systems to Ni-Cr alloy)

  • 송소연;손병화;김종엽;신상완;이정열
    • 대한치과보철학회지
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    • 제53권4호
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    • pp.295-300
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    • 2015
  • 목적: 레진과 니켈-크롬 합금의 결합력에 미치는 금속 프라이머 및 다용도 접착제의 영향에 대하여 비교 평가하고자 한다. 재료 및 방법: 실험을 위해 120개의 니켈-크롬 합금(Vera Bond 2V) 디스크를 제작하여 아크릴 레진 실린더에 포매하였다. 시편의 표면은 220 grit, 600 grit의 실리콘 카바이드지로 연마한 뒤 $50{\mu}m$의 알루미늄옥사이드 입자를 분사하여 처리하였다. 실험군은 메탈 프라이머의 적용(Metal primer II, Alloy primer, Metal & Zirconia primer, MKZ primer)과 다용도 접착 시스템(Single Bond Universal, All Bond Universal)에 따라 6개 군으로 나뉘었다. 각 시편의 중앙에 높이 2 mm, 직경 3 mm로 복합레진을 충전하였으며, 제작된 모든 시편은 $37^{\circ}C$ 증류수에서 24시간 보관하였다. 만능시험기에 시편을 위치시킨 후 1 mm/min cross head speed로 전단결합강도를 측정하였다. 통계분석은0.05의 유의수준으로 일원분산분석을 시행하였고 Tukey's multiple coMParison test로 사후검정을 하였다. 결과: Single Bond Universal, All Bond Universal, Metal Primer II 3개 군과 Alloy Primer, MKZ Primer, Metal & Zirconia Primer 3개 군 사이에 통계학적으로 유의한 차이를 보였다(P<.001). 결론: 니켈-크롬 합금에 대한 다용도 접착 시스템의 전단결합강도는 Metal Primer II를 제외한 기존의 금속 프라이머를 적용했을 때 보다 높게 나타났다. 본 실험에서 더 나아가 다용도 접착 시스템 내 실란(Silane) 포함 여부에 따른 효과를 평가할 수 있는 연구가 필요할 것이다.

일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용 (Application for Identification of Food Raw Materials by PCR using Universal Primer)

  • 박용춘;진상욱;임지영;김규헌;이재황;조태용;이화정;한상배;이상재;이광호;윤혜성
    • 한국식품위생안전성학회지
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    • 제27권3호
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    • pp.317-324
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    • 2012
  • 본 연구는 식품원료의 진위여부를 판별하기 위한 시험법으로 일반 프라이머를 이용한 DNA barcode 기법을 도입하였다. 동물성식품원료의 경우 미토콘드리아 DNA 중 cytochrome oxidase subunit I(COI) 부위 검출을 위하여 디자인된 프라이머(LCO1490/HCO2198 및 VF2/FISH R2)와 cytochrome b(cyt b) 부위 검출을 위하여 디자인된 프라이머(L14724/H15915)를 사용하였다. 상기 3 종류의 프라이머를 사용하여 가축류 6종(소, 돼지, 염소, 양, 말 및 사슴), 가금류 4종(닭, 오리, 칠면조 및 타조), 어류 7종(명태, 대구, 청대구, 청어, 송어, 다랑어 및 우럭)을 대상으로 PCR 후 전기영동하여 예상되는 PCR 산물의 생성 유무를 확인하였다. 가축류 6종에 대하여는 LCO1490/HCO2198, VF2/FISH R2 및 L14724/H15915 프라이머를 사용한 경우 COI 및 cyt b가 모두 검출되었으며, 가금류 4종은 LCO1490/HCO2198 및 VF2/FISH R2 프라이머를 사용한 경우만 COI이 검출되었다. 또한 어류 7종은 VF2/FISH R2 프라이머를 사용한 경우에만 COI 부위가 검출됨을 확인하였다. 식물의 경우 엽록체 DNA 부위를 이용하여 디자인된 3 종류의 프라이머(trnH/psbA, rpoB 1F/4R 및 rbcL 1F/724R)를 사용하였다. 각각의 프라이머를 이용하여 식물 5종(마늘, 양파, 무, 녹차 및 시금치)에 대하여 실험한 결과 3종류의 프라이머에서 PCR의 산물을 모두 확인하였으며 trnH/psbA 프라이머의 경우 식물 종마다 PCR 산물의 크기는 다르게 검출되었다. 본 연구에서는 17종의 식품원료별 일반 프라이머 및 PCR 조건을 확립하였으며, 생산된 PCR 산물을 대상으로 염기서열을 결정하고 유전자은행에 있는 염기서열과 DB 비교 분석을 통하여 식품에 사용된 원료의 진위여부 판별에 적용이 가능할 것으로 기대된다.

Universal Rice Primer (URP)-RAPD 방법에 의한 어류 종 특이 marker의 동정 (Identification of Potential Species-Specific Marker in Several Fish Species by RAPD Using Universal Rice Primers)

  • 김우진;김경길;이정호;박두원
    • 한국수산과학회지
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    • 제36권3호
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    • pp.317-320
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    • 2003
  • Morphologically similar fish species were subjected to the random amplified polymorphic DNA (RAPD) analysis using universal rice primer (URP). The fish species tested were sea basses (Lateolabrax japonicus and L. maculatus), eels (Anguilla japonica, A. bicolor bicolor, A. rostrata, and A. anguilla), and flounders (Limanda yokohamae and L. herzensteinin). Highly reproducible RAPD patterns were observed with several potential species-specific markers. The results indicate that RAPD technique using URP is useful for distinguishing fish psecies in a rapid manner.

Genetic Variation in Geographic Crayfish (Cambaroides similis) Populations

  • 윤종만;김용호;김솔
    • 한국어병학회지
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    • 제19권2호
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    • pp.141-153
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    • 2006
  • Genomic DNA samples isolated from two geographical crayfish (Cambaroides similis) populations in the inland of the Korean Peninsula, at Jeonju (Jeonju crayfish; JJC) and Jeongup (Jeongup crayfish; JUe), were PCR-amplified repeatedly. The six arbitrarily selected primers OPC-03, OPC-06, OPC-09, URP-02, URP07 and URP-09 generated the common, specific, and polymorphic fragments. The sizes of DNA fragments also varied widely, from 100 bp - 2,600 bp. Here, 521 fragments were identified in the JJC population, and 354 in the JUC population: 6 primers generated 60 specific fragments (60/521 fragment, 11.5%) in the JJC population, and 90 (90/354 fragments, 25.4%) in the JUC population. These primers produced 42 polymorphic fragments (8.1%) in the DC population, and 18 (5.1%) in the mc population. Especially these results demonstrate that the primers detected numerous specific fragments. Especially, the decamer primer OPC-06 generated inter-population-common DNA fragments, approximately 400 and 800 bp, respectively, in both the JJC and JUC populations. The universal primer URP-02 also generated inter-population-identical DNA fragments, approximately 350 bp and 600 bp, between the two geographical crayfish populations. Based on the average bandsharing values of all samples, the bandsharing value of individuals within the JJC population was much higher than in the JUC population. The bandsharing value between individuals no. 10 and no. 15 was 0.683, which was the highest between the two geographical populations. The dendrogram obtained by the six primers indicates two genetic clusters: cluster I (CRAYFISH 01 - CRAYFISH II), and cluster 2 (CRAYFISH 12 - CRAYFISH 22). The genetic distance between the two geographical populations ranged from 0.053 to 0.605. Ultimately, the longest genetic distance displaying significant molecular differences was found to exist between individuals in the two crayfish populations, between individuals CRAYFISH no. 02 of Jeonju and CRAYFTSH no. 15 of Jeongup (genetic distance = 0.605).

Effect of adhesive primers on bonding strength of heat cure denture base resin to cast titanium and cobalt-chromium alloy

  • Kim, Su-Sung;Vang, Mong-Sook;Yang, Hong-So;Park, Sang-Won;Lim, Hyun-Pil
    • The Journal of Advanced Prosthodontics
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    • 제1권1호
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    • pp.41-46
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    • 2009
  • STATEMENT OF PROBLEM. The poor chemical bonding of a denture base resin to cast titanium framework often introduces adhesive failure and increases microleakage. PURPOSE. This study evaluated the shear bond strengths of a heat cure denture base resin to commercially pure titanium, Ti-6Al-4V alloy and a cobalt-chromium alloy using two adhesive primers. MATERIAL AND MATHODS. Disks of commercially pure titanium, Ti-6Al-4V alloy and a cobalt-chromium alloy were cast. Specimens without the primer were also prepared and used as the controls. The shear bond strengths were measured on a screw-driven universal testing machine. RESULTS. The primers significantly(P < .05) improved the shear bond strengths of the heat cure resin to all metals. However, the specimens primed with the Alloy $primer^{(R)}$(MDP monomer) showed higher bond strength than those primed with the MR $bond^{(R)}$(MAC-10 monomer) on titanium. Only adhesive failure was observed at the metal-resin interface in the non-primed specimens, while the primed specimens showed mixed failure of adhesive and cohesive failure. CONCLUSIONS. The use of appropriate adhesive metal primers makes it possible not only to eliminate the need for surface preparation of the metal framework before applying the heat cure resins, but also reduce the need for retentive devices on the metal substructure. In particular, the Alloy $primer^{(R)}$, which contains the phosphoric acid monomer, MDP, might be clinically more acceptable for bonding a heat cure resin to titanium than a MR $bond^{(R)}$, which contains the carboxylic acid monomer, MAC-10.

Restriction Analyses of PCR Amplified Partial SSU Ribosomal DNA to Distinguish Arbuscular Mycorrhizal Fungi from Other Fungi Colonizing Plant Roots

  • Lee, Jae-Koo;Tae, Moon-Sung;Eom, Ahn-Heum;Lee, Sang-Sun
    • Mycobiology
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    • 제31권2호
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    • pp.68-73
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    • 2003
  • Roots of Glycine max and Miscanthus sinensis and soil samples were collected from various field sites at Goesan, Chungbuk in Korea. Microscopic observations of the roots indicated high colonization rates of both arbuscular mycorrhizal fungi(AMF) and other fungi. The partial small subunit of ribosomal DNA genes were amplified with the genomic DNA extracted from their roots by nested polymerase chain reaction(PCR) with universal primer NS1 and fungal specific primers AML Restriction fragment length polymorphism(RFLP) was analyzed using the combinations of three restriction enzymes, HinfI, AluI and AsuC21. Nucleotides sequence analysis revealed that ten sequences from Miscanthus sinensis and one sequence from Glycine max were close to those of arbuscular mycorrhizal fungi. Also, 33% of total clones amplified with NS31-AM1 primers from M. sinensis and 97% from G. max were close to Fusarium oxysporum or other pathogenic fungi, and they were successfully distinguished from AME Results suggested that these techniques could help to distinguish arbuscular mycorrhizal fungi from root pathogenic fungi in the plant roots. Especially, DNA amplified by these primers showed distinct polymorphisms between AMF and plant pathogenic species of Fusarium when digested with AsuC21.

Thermal Cycling에 따른 자가중합 레진의 결합강도에 관한 연구 (THE EFFECTS OF THERMAL CYCLING ON THE BOND STRENGTH OF SELF-CURING RESIN)

  • 조혜원;하점임
    • 대한치과보철학회지
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    • 제35권4호
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    • pp.697-705
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    • 1997
  • The purpose of this study was to evaluate the effects of two metal adhesive primers on the shear bond strengths of self-curing resin to Ni-Cr a]toy and the effects of 1000 thermal cycling on the durability of the bond. The two selected metal adhesive primers were Metal Primer II(G-C corp., Japan) and MR Bond(Tokuyama corp., Japan) and no treatment groups were used as control. All specimens were divided into two groups according to thermal cycling. In the group without thermal cycling, the specimens were stored in water for 24 hours. In the group with thermal cycling, the specimens were thermocycled 1000 times at temperature of $5^{\circ}C\;and\;55^{\circ}C$. Shear bond strengths were measured using the Universal testing machine(Zwick 145641, Germany) with a crosshead speed of 0.5 mm/min. The results were as follows: 1. MR Bond significantly improved the shear bond strength of resin to Ni-Cr alloy before and after thermal cycling. 2. There were no difference in the shear bond strength of resin to Ni-Cr alloy between Metal Primer II treated group and no treatment group. 3. Regardless of the type and the use of adhesive primers, there were tendency of decrease in shear bond strength with 1000 thermal cycling.

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Development of Clamping Probe for Rare DNA Detection using Universal Primers

  • Kim, Meyong Il;Lee, Ki-Young;Cho, Sang-Man
    • Fisheries and Aquatic Sciences
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    • 제17권3호
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    • pp.339-344
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    • 2014
  • PCR amplification with universal primer is a useful tool for speciation of symbionts in marine eukaryote coupled with robust separation method such as denaturing high performance chromatography (DHPLC). To overcome the biased amplification, clamping PCR is recommended to suppress the amplification of host gene. In this study, we evaluated the efficiency of rare gene detection for two kinds of clamping probes which were successfully utilized for eukaryotic symbiont analysis: C3 linked nucleotide (C3) and peptide nucleic acid (PNA). PNA was 3-4 orders of magnitude higher than that of C3 tested in clamping efficiency and rare gene detection. This represented that PNA could be a more competent clamping probe for the enhancement of PCR amplification for rare symbiont genes.