• Korean Journal of Plant Resources
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• v.17 no.2
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• pp.107-115
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• 2004

To develop universal primers for phylogenetic analysis and species-specific marker for breeding program of kiwifruit, eighteens primers were designed from kiwifruit genome-specific repeat sequences. Seven species including twenty two varieties collected from native eastern Asia were examined using 18 to 22 mer kiwifruit target(KT) primers. among eighteen primers, we selected seven primers for phylogenetic relationship. The genus Actinidia was divided into two large groups; group I,A. arguta, A. melanandra, A. kolomikta, and A. marcrosperma, characterized by the non-hair in fruits and loaves or a few pubescences only in young stage, which belongs to the section Leiocarpae, and group II, A. chinensis, A. deliciosa, and A. eriantha, characterized by a lot of hairs only in young fruit stage and with a lot of hairs or fuzzes in leaves and branches, which belongs to the section Stellatae. Group II especially belongs to the series Perfectae of the section Stellatae and was divided into two subgroups; subgroup I containing A. chinensis and A. deliciosa, and subgroup II containing A. eriantha. In contrast, the two species, A. chinensis and A. deliciosa, which are known to have common parents, were divided into two independent subgroups with 80％ of a similarity value. On the other hand, we selected KT6F for variety specific bands, KT12E primers for 'Hayward' and 'Tomuri'. KT7F or KT12F primers were useful for analysis of inheritance pattern in kiwifruit cross-breeding. We suggest that these primers will be a powerful tool for elucidating phylogenetic relationship and selection of novelty kiwifruit in a breeding program.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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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.

• The Journal of Korean Academy of Prosthodontics
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• v.53 no.4
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• pp.295-300
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• 2015

Purpose: The aim of this study was to evaluate the shear bond strength between Ni-Cr alloy and composite resin using universal adhesive systems coMPared to conventional method using metal primers. Materials and methods: For this study, a total of 120 cast commercial Ni-Cr alloy (Vera Bond 2V) disks were embedded in acrylic resin, and their surfaces were smoothed with silicon carbide papers and airborne-particle abrasion. Specimens of each metal were divided into 6 groups based on the combination of metal primers (Metal primer II, Alloy primer, Metal & Zirconia primer, MKZ primer) and universal adhesive systems (Single Bond Universal, All Bond Universal). All specimens were stored in distilled water at $37^{\circ}C$ for 24 hours. Shear bond strength testing was performed with a universal testing machine at a cross head speed of 1 m/min. Data (MPa) were analyzed using one-way ANOVA and the post hoc Tukey's multiple comparison test (${\alpha}$=.05). Results: There were significant differences between Single Bond Universal, All Bond Universal, Metal Primer II and Alloy Primer, MKZ Primer, Metal & Zirconia Primer (P<.001). Conclusion: Universal Adhesive system groups indicated high shear bond strength value bonded to Ni-Cr alloy than that of conventional system groups using primers except Metal Primer II. Within the limitations of this study, improvement of universal adhesive systems which can be applied to all types of restorations is recommended especially non-precious metal alloy. More research is needed to evaluate the effect of silane inclusion or exclusion in universal adhesive systems.

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.317-324
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• 2012

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.3
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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.

• Journal of fish pathology
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• v.19 no.2
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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).

• The Journal of Advanced Prosthodontics
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• v.1 no.1
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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.

• Mycobiology
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• v.31 no.2
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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.

• The Journal of Korean Academy of Prosthodontics
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• v.35 no.4
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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.

• Fisheries and Aquatic Sciences
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• v.17 no.3
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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.