• Title/Summary/Keyword: PCR technique

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Identification of Meat Species Using Species-Specific PCR-RFLP Fingerprint of Mitochondrial 12S rRNA Gene (미토콘드리아 12S rRNA 유전자의 종 특이적 PCR-RFLP Fingerprint를 이용한 식육 원료의 판별)

  • Park, Jong-Keun;Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.27 no.2
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    • pp.209-215
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    • 2007
  • In order to develop a sensitive and reliable method for the species-specific molecular markers, PCR-RFLP assay of the mitochondrial DNA(mt DNA) 12S rRNA gene was exploited for the identification of the origin of animal meat species including cattle, pig, sheep, goat, horse, deer, chicken, duck and turkey. A specific primer pairs were designed, based on the nucleotide sequences of mt 12S rRNA gene, for the amplification of the highly conserved region in the gene of the animal species using PCR-RFLP technique. mt DNA was isolated from meat samples followed by DNA amplification using PCR with the specific primers. PCR amplification produced an approximately 455 bp fragment in each of these animal meats. To distinguish pleat species, the PCR amplicons were digested with restriction endonucleases Tsp5091 and MboI, respectively, which generates distinct RFLP profiles. The DNA profiles digested with Tsp5091 allowed the clear discrimination in the mammalian meat species and the DNA profiles digested with MboI in poultry meat species. Therefore, the PCR-RFLP profiles of mt 12S rRNA gene could be very useful to identify the origin of the raw materials in the raw meats as well as the processed meat products.

Identification of differentially expressed genes using an annealing control primer system in periodontitis

  • Na, Hee-Sam;Kim, Ji-S.;Chung, Jin
    • International Journal of Oral Biology
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    • v.37 no.3
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    • pp.109-114
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    • 2012
  • In the gingival tissues of patients with periodontitis, inflammatory responses are mediated by a wide variety of genes. In this study, we screened for differentially expressed genes (DEGs) in periodontitis compared with normal tissue using an annealing control primer (ACP) system. By ACP RT-PCR analysis, we obtained about 160 amplicons, 8 of which were found to be differentially expressed. DEGs in patients with periodontitis were thus successfully and reliably identified by the ACP-based RT PCR technique. The DEGs identified in the screen may also enhance our understanding of the pathogenesis of periodontitis.

Analysis of Mitochondrial DNA Mutation in hepatoma

  • Chung, Ku-Sun;Lee, Kyo-Young;Shim, Sang-In;Kim, Jin-Sun;Song, Eun-Sook
    • BMB Reports
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    • v.33 no.5
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    • pp.417-421
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    • 2000
  • Mitochondrial DNA (mtDNA) mutation was investigated in a hepatoma patient using a polymerase chain reaction (PCR) and an in situ hybridization technique. Biotin-labeled probes for the subunit m of cytochrome c oxidase revealed differences in the in situ hybridization. A PCR assay using biopsied and microdissected tissues showed that common deletion (4,977 bp) was more pronounced in the cancer region than in the normal parts of the same patient. These results suggest that mtDNA deletion might be associated with tumorigenesis in hepatoma.

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A Method for Evaluation of the Quality of DNA Microarray Spots

  • Zhang, Bao;Ma, Wen-Li;Hu, Zi-You;Shi, Rong;Song, Yan-Bin;Zheng, Wen-Ling
    • BMB Reports
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    • v.35 no.5
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    • pp.532-535
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    • 2002
  • To establish a method to evaluate the quality of the printed microarray and DNA fragments' immobilization. The target gene fragments that were made with the restriction display PCR (RD-PCR) technique were printed on a superamine modified glass slide, then immobilized with UV cross-linking and heat. This chip was hybridized with universal primers that were labeled with cy3-dUTP, as well as cDNA that was labeled with cy3-dCTP, as the conventional protocol. Most of the target gene fragments on the chip showed positive signals, but the negative control showed no signal, and vice versa. We established a method that enables an effective evaluation of the quality of the microarrays.

Studies on the Endocrine Disruption in Wildlife Fish (어류생체지표를 이용한 내분비계장애 연구)

  • 구자민;류지성;정규혁;이철우;박응로;박광식
    • Environmental Analysis Health and Toxicology
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    • v.16 no.4
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    • pp.197-204
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    • 2001
  • Endocrine disruption in crucian carp (Carassius auratus) living in the branch of Han River were examined. Vitellogenin level in plasma was measured using ELISA system and aromatase mRNA level in brain was observed using RT-PCR technique. In all female fish, vitellogenin levels were in the range of 20∼40 $\mu\textrm{g}$/ml and aromatase mRNA expression could be detected on the agarose gel after RT-PCR. However, in case of males, vitellogenin level was elevated in only one fish, while vitellogenin was hardly detected in others. Aromatase was expressed in all males although the levels were relatively lower than the level in female fish. Testis-ova and any other histological changes of reproductive organ were not shown in both sexes.

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Multiplex PCR Technique for Rapid Detecting of Hanwoo and Holstain (한우 및 젖소를 고속으로 판별하기 위한 멀티 플레스 PCR 기술 개발)

  • Lee, Jin-Sung
    • Proceedings of the KAIS Fall Conference
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    • 2007.05a
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    • pp.46-47
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    • 2007
  • 본 논문은 소의 모색에 관여하는 Melanocortin I recepter (MCIR) 유전자의 염기서열을 기초로 정확하게 한우와 젖소를 구분할 수 있는 단일 염기 변이를 이용하여 기존의 방법보다 2배 이상으로 빠른 속도로 한우와 젖소를 판별할 수 있는 multiplex PCR 기술에 관한 것이다. 또한, 동 기술에 적용된 기술에는 기존의 방법으로는 동시 판별을 할 수 없었던 소의 암수 구분이 가능하도록 수컷 판별을 위한 표지 유전자인 SRY 유전자를 적용함으로써 한우와 젖소 그리고 압수를 고속으로 동시 판별이 가능하게 되어 젖소가 한우로 둔갑되는 현재의 불법 유통의 문제점을 해결해 주는 새로운 방법이 될 수 있을 것으로 사료된다.

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Secretion of escherichia coli $\beta$-lactamase from bacillus subtilis with the aid of usufully constructed secretion vector

  • Park, Geon-Tae;Rho, Hyun-Mo
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.60-64
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    • 1992
  • The secretion vector with promoter and signal sequence region of neutral protease gene (npr) from Bacillus amyloliquefaciens was constructed by the technique of polymerase chain reaction (PCR). A unique restriction iste was introduced into the 3' of the signal coding region by the synthesis of PCR primer. To demonstrate the function of cloned promoter and signal sequence, we used the E. coli .betha.-lactamase structural gene as a foreign gene. The signal sequence of .betha.-lactamase gene was deleted by Bal31 exonuclease and only mature region was introduced into the secretion vector. Bacillus subtilis cells transformed by the recombinant vector synthesized the fusion protein and were also capable of removing the signal peptide from the original fusion protein, as judged by the assay of .betha.-lactamase activity and secretion into the growth medium by western blotting.

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Vitellogenin mRNA Induction in Male Carp Treated with Bisphenol A (비스페놀A 처리 수컷 잉어에서 비텔로제닌 mRNA의 발현)

  • 박응로;최필선;이철우;류지성;남성숙;류홍일;전성환;나진균;박광식
    • Toxicological Research
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    • v.17 no.3
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    • pp.181-186
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    • 2001
  • The estrogenic potency of bisphenol A using reverse trancriptase-PCR response of liver vitellogenin mRNA in male carp was studied. For this, six combination of primers which were synthesized on the basis of cDNA consensus region of various species, were evaluated and one pair of primers was selected as the best to show 286 bp size-transcript. By using the selected primers, vitellogenin mRNA induction in carp treated with bisphenol A was measured and the chemical showed dose-and time-dependent Induction response. From this result, it was concluded that RT-PCR technique wing the selected primers in this study can be wed to monitor the estrogenic effects exerted In carp living in Korean freshwater.

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Rapid Identification of Potato Scab Causing Streptomyces spp. Using Pathogenicity Specific Primers

  • Shin, Pyung-Gyun;Kim, Jeom-Soon;Hahm, Young-Il
    • The Plant Pathology Journal
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    • v.18 no.6
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    • pp.338-341
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    • 2002
  • The potato scab is caused by several species of Streptomyces. Among these species, only pathogenic strains were found to produce thaxtomin A characterized by necrotic bioassay and HPLC. In this study, identification of the pathogenic strains of Streptomyces was performed through the polymerase chain reaction (PCR) by using specific pathogenicity primer sets derived from the nec1 gene sequences of Streptomyces scabies. The expected PCR products were obtained approximately 580 bp and confirmed by sequencing. This PCR technique can be used effectively to identify the pathogenic Streptomyces species, that cause scab on potato tubers.

Analysis of Geographical Genetic Differences of Arkshell Populations in Korea

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.25 no.2
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    • pp.105-111
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    • 2021
  • A PCR-founded genetic analysis aim and principle was used to foster a hierarchical polar dendrogram of the Euclidean genetic distances (GDs) for two arkshell populations, Scapharca broughtonii (YEOSU, Yeosu population and JINHAE, Jinhae population). Five oligonucleotides primers were make use of to craft 354 and 390 scorable bands in the Yeosu and Jinhae populations, respectively, outspreading in DNA fragment size from 100 bp to 1,600 bp. The bandsharing (BS) results disclosed that the Jinhae population had a higher average BS value (0.700) than that for the Yeosu population (0.692). The GD between individuals supported an adjacent association in grouping II (JINHAE 12 - JINHAE 22). The observation of a noteworthy GD between the two Scapharca populations verified that this PCR-generated technique could be a profitable attempt for within- and between-population-grounded biological DNA scrutiny. The potential of PCR inquiry will be favorable in the selection of individuals and/or populations for several reproductive- and/or quarantine-connected characters in aquafarming manufacture.