• Title/Summary/Keyword: primer coat

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Detection of Lily Symptomless Virus Using RT-PCR Technique (RT-PCR 기법을 이용한 Lily Symptomless Virus의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.187-190
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    • 1996
  • 백합으로부터 total RNA를 분리하여 LSV 외피단백질 유전자의 551 bp에 해당하는 특정 염기서열을 증폭할 수 있는 primer로 RT-PCR를 수행하였다. 그 결과 Lilium oriental hybrid cvs. Miani, Marco Polo, Casablanca, Le Reve 품종에서 551 bp의 DNA 절편이 증폭되었고 이 절편의 염기서열을 분석한 결과 LSV외피단백질 유전자의 일부임을 확인할 수 있었다. 그러므로 RT-PCR 방법으로, 실험에 사용하 s4품종 모두 LSV에 감염되어 있음을 알 수 있었다.

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Corrosion Protection of Automotive Steels by Novel Water-borne Primer Systems

  • Ooij, William J. van;Puomi, Paula
    • Corrosion Science and Technology
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    • v.6 no.5
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    • pp.239-244
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    • 2007
  • Corrosion protection of automotive steels has traditionally been assured by using a zinc phosphate metal pretreatment followed by the deposition of a cathodic electrocoat system. This system has been developed and optimized over the years into a highly robust and dependable system with a high performance. However, in terms of efficiency and use of resources and energy, the need is now felt to develop a simpler system with fewer steps, shorter lines, less energy requirements (curing and e-coat deposition) and less stringent waste disposal requirement (phosphate sludge). We report here on the development of a one-step system that can possibly replace both the zinc phosphate and the e-coating processes. Such a system is based on the so-called superprimer concept that we have recently developed for the replacement of chromate pretreatment and chromate-containing primers in the aerospace industry. With some modifications, such systems can also be adapted for use in the automotive industry.

Application of High Performance Coatings for Service Life Extension of Steel Bridge Coatings

  • Lee, Chan-Young
    • Corrosion Science and Technology
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    • v.20 no.4
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    • pp.169-174
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    • 2021
  • In this study, performance tests, a field evaluation, and a life cycle cost (LCC) analysis for high performance coating systems were conducted to prepare a plan to reduce the cost of maintenance coating and contribute to the service life extension of steel bridges by applying high performance coatings to steel bridges that will be constructed in the future. From the deterioration models based on the field evaluation for chlorinated rubber and urethane topcoat systems, which have been applied often, the mean service lives were derived as 20.8 and 26.6 years, respectively. For the other coating systems that have not been applied in practice, the coordination factors were differentially applied with evaluation items. The most durable coating system was predicted to be thermal spray coating (TSC) primer/epoxy intermediate coat/fluoride resin topcoat, with a predicted value as long as 42.2 years. The LCC analysis indicates that partial application of high performance coating, such as TSC and fluoride resin, to specific parts vulnerable to corrosion and ultraviolet ray (UV) is more advantageous than the use of general coating systems.

분자유전학적인 기술을 이용한 육 감별법

  • Kim, Tae-Heon
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2000.11a
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    • pp.59-75
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    • 2000
  • This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

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Bond strength of different bonding systems to the lingual surface enamel of mandibular incisors (하악 전치 설측면에 대한 다양한 접착시스템의 접착강도)

  • Turkoz, Cagri;Tuncer, Burcu Balos;Ulusoy, Mehmet Cagri;Tuncer, Cumhur
    • The korean journal of orthodontics
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    • v.40 no.4
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    • pp.260-266
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    • 2010
  • Objective: The aim of this study was to determine whether different types of adhesive systems and enamel-protective agents will affect the tensile bond strength of lingual brackets. Methods: A total of 75 extracted mandibular incisors were randomly divided into 5 groups and lingual brackets were bonded. Group 1 specimens received Transbond XT (3M Unitek, Monrovia, CA, USA), Group 2 required the application of a fluoride-releasing resin (Ortho-coat, Pulpdent, Watertown, MA, USA) with Transbond XT, Group 3 specimens received a chlorhexidine varnish (Cervitec Plus, Ivoclar Vivadent, Schaan, Lichtenstein) with Transbond XT. In Group 4, a light-cured orthodontic adhesive (Aegis Ortho, Bosworth, Skokie, USA) was applied and in Group 5, an antimicrobial self-etching primer (Clearfil Protect Bond, Kuraray, Osaka, Japan) was used. Results: There were no significant differences in bond strength whether fluoride-releasing resin or chlorhexidine varnish were used or not. Group 5 had significantly higher bond strength and adhesive remnant index (ARI) values than other groups (p < 0.001). The application of enamel-protective products did not have an adverse affect on the bond strength of lingual brackets. Conclusions: These products might provide benefits both for the patient and the clinician, by supporting the oral hygiene during lingual orthodontic treatment. The higher ARI score may be beneficial for Clearfil Protect Bond but its excessive bond strength should be considered in clinical practice, especially where the enamel is thin.

Characterization of Cucumver mosaic virus Isolated from Hydrangea macrophylla for. otaksa (Sieb. et Zucc) Wils. (수국에서 분리한 Cucumber mosaic virus의 특성)

  • 방주희;박선정;이금희;최장경;이상용
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.1-7
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    • 2001
  • An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

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Isolation and Characterization of Watermelon Isolate of Cucumber green mottle mosaic virus(CGMMV-HY1) from Watermelon Plants with Severe Mottle Mosaic Symptoms

  • Shim, Chang-Ki;Han, Ki-Soo;Lee, Jung-Han;Bae, Dong-Won;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.167-171
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    • 2005
  • We isolated the Cucumber green mottle mosaic virus(CGMMV) particles from watermelon leaves and designated as CGMMV-HY1 as a watermelon isolate and attempted to characterize the pathogenic isolate responsible for such an epidemic in watermelon and also to monitor dominant viral isolates in greenhouse. The watermelon plants infected with CGMMV generally showed mottle mosaic, mosaic, growth stunting, necrosis and deformed fruit. The reactions of indicator plants to CGMMV-HY1 were the local lesions on Nicotiana tabacum cv. White Burley, Nicotiana tabacum cv. Samsun, and Chenopodium amaranticola, and the mosaic symptoms only on Cucumis sativus, but the CGMMV-HY1 did not infect Nicotiana sylvesytis, Datura stramonium, Chenopodium quinoa, and Petunia hybrida. Purified virus particles were rod-shaped and about 300 nm long. The coat protein (CP) of purified CGMMV-HY1 was single band with molecular weight of about 16.5 kDa which was confirmed by western blot analysis probed with monoclonal antibody of CGMMV-HY1. The genomic and subgenomic RNAs of 6.4 kb and 0.75 kb were revealed by the electrophoresis on 1.2% formaldehydedenatured agarose gel. Viral and complementary CGMMV-specific primer sets were designed for spanning the genome using previously reported CGMMV sequences. A 464bp of CP gene of CGMMV-HY1 was amplified by RT-PCR and cloned into PGEM-T easy vector. The nucleotide sequence of CP gene of CGMMV-HY1 shared 98%, 99%, and 100% identities with that of CGMMV strains W, KOM, and KW respectively. Based on these results, we identified CGMMV-HY1 as a CGMMV isolate of watermelon, a member of Tobamovirus.

Characterization of Peanut stunt virus Isolated from Black Locust Tree (Robinia pseudo-acacia L.)

  • Bang, Ju-Hee;Choi, Jang-Kyung;Lee, Sang-Yong
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.125-130
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    • 2006
  • An isolate of Peanut stunt virus (PSV) isolated from black locust tree (Robinia pseudo-acacia L.) showing severe mosaic and malformation symptoms, was designated as PSV-Rp. PSV-Rp was characterized by the tests of host range, physical properties, RNA and coat protein composition and RT-PCR analysis. Nucleotide sequences of the cucumoviruses CP genes were also used for identification and differentiation of PSV-Rp. Six plant species were used in the host range test of PSV-Rp. PSV-Rp could be differentiated from each Cucumovirus strain used as a control by symptoms of the plants. The physical properties of PSV-Rp virus were TIP $65^{\circ}C$, DEP $10^{-3}$, and LIP $2{\sim}3$ days. In dsRNA analysis, PSV-Rp consisted of four dsRNAs, but satellite RNA was not detected. Analysis of the coat proteins by SDS-PAGE showed one major protein band of about 31 kDa. RT-PCR using a part of Cucumovirus RNA3 specific primer amplified ${\sim}950bp$ DNA fragments from the crude sap of virus-infected black locust leaves. RFLP analysis of the RT-PCR product could differential PSV-RP from CMV The nucleotide sequence identity between the PSV-Rp CP and the TAV-P CP genes and the PS-V-RP CP and CMV-Y CP genes were 61.6% and 40.5%, respectively. On the other hand, the nucleotide sequence identity of the PSV-Rp CP gene was $70.9%{\sim}73.4%$ in comparison with those of PSV subgroup I (PSV-ER and PSV-J) and 67.3% with that of PSV subgroup II(PSV-W). Especially, the nucleotide sequence identity of PSV-Rp CP gene and that of PSV-Mi that was proposed recently as the type member of a novel PSV subgroup III was 92.4%.

Biological, Physical and Cytological Properties of Pepper mottle virus-SNU1 and Its RT-PCR Detection

  • Han, Jung-Heon;Choi, Hong-Soo;Kim, Dong-Hwan;Lee, Hung-Rul;Kim, Byung-Dong
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.155-160
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    • 2006
  • A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

Characterization of Tobacco rattle virus(TRV-K) isolated in Korea (한국에서 분리한 Tobacco rattle virus(TRV-K)의 특성)

  • Shin, Hye-Houng;Koo, Bong-Jin;Kang, Sang-Gu;Chang, Moo-Ung;Ryu, Ki-Hyung
    • Research in Plant Disease
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    • v.8 no.4
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    • pp.207-214
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    • 2002
  • Tobacco rattle virus(TRV) was detected from Gladiolus hybridus, Crocus spp. and Narcissus spp. leaves show-ing notched or stripe on the leaf and malformation symptoms collected from Daegu and Kyungbuk province by electron microscopy (EM), immunosorbent electron microscopy (ISEM) and host range study. Direct negative staining method by EM showed rigid rod long particles 170~200$\times$22 nm and rigid rod short particles 40~114$\times$22 m. TRV-K isolated from G. hybridus propagated with Nicotiana tabacum. TRV coat protein(CP) gene was amplified using specific oligonucleotide primer by RT-PCR. Sequence analysis of amplified CP gene showed 99.5% nucleotide similarity to TRV-ORY.