• 제목/요약/키워드: Microsatellite DNA

검색결과 259건 처리시간 0.031초

Retrotransposon Microsatellite Amplified Polymorphism Strain Fingerprinting Markers Applicable to Various Mushroom Species

  • Le, Quy Vang;Won, Hyo-Kyung;Lee, Tae-Soo;Lee, Chang-Yun;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
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    • 제36권3호
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    • pp.161-166
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    • 2008
  • The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with $\underline{Re}trotransposon$ $\underline{M}icrosatellite$ $\underline{A}mplified$ $\underline{P}olymorphism$ (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.

한국 토종오리의 개체 식별 및 품종 구분을 위한 Microsatellite 마커 탐색 (Investigation of Microsatellite Markers for Traceability and Individual Discrimination of Korean Native Ducks)

  • 서동원;술타나;최누리;김연수;진실;허강녕;진선덕;이준헌
    • 한국가금학회지
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    • 제42권1호
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    • pp.1-8
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    • 2015
  • 최근, 한국의 소비자들이 건강에 대한 관심이 증가하면서 단일불포화 지방산이 풍부해 건강에 긍정적인 영향을 줄 수 있는 오리고기의 수요가 급격하게 증가하고 있다. 하지만 대부분의 종 오리는 수입에 의존하고 있는 실정이기 때문에, 토종오리의 개발 및 보급이 필요한 실정이며, 이는 종자주권의 확립 및 농가 소득 증대에도 매우 필요한 일이라 할 수 있다. 따라서 본 연구에서는 24개의 microsatellite 마커를 확보하였으며, 이들 마커의 대립유전자수는 1~16개, 이형접합도는 0~0.865, 다형성은 0~0.841로 확인되었다. 이들 마커를 이용하여 임의 집단에서 동일개체 출현빈도를 계산한 결과는 임의 집단 $1.64{\times}10^{-16}$, 전형매 집단 $2.60{\times}10^{-7}$, 반형매 집단 $1.30{\times}10^{-12}$으로 높은 개체식별률과 친자확인도를 확인할 수 있었다. 하지만 이들 마커를 이용한 계통분석 결과, 토종오리와 실용오리 집단을 정확하게 구분하기에는 어려운 것으로 확인되었다. 따라서 추가연구를 통해 토종오리의 순종화 및 더 정확한 토종오리와 실용오리 집단 구분이 가능한 마커 개발이 필요할 것으로 사료된다.

Inter Simple Sequence Repeat (ISSR) Polymorphism and Its Application in Mulberry Genome Analysis

  • Vijayan Kunjupillai
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권2호
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    • pp.79-86
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    • 2005
  • Molecular markers have increasingly been used in plant genetic analysis, due to their obvious advantages over conventional phenotypic markers, as they are highly polymorphic, more in number, stable across different developmental stages, neutral to selection and least influenced by environmental factors. Among the PCR based marker techniques, ISSR is one of the simplest and widely used techniques, which involves amplification of DNA segment present at an amplifiable distance in between two identical microsatellite repeat regions oriented in opposite direction. Though ISSR markers are dominant like RAPD, they are more stable and reproducible. Because of these properties ISSR markers have recently been found using extensively for finger printing, pohylogenetic analysis, population structure analysis, varietal/line identification, genetic mapping, marker-assisted selection, etc. In mulberry (Morus spp.), ISSR markers were used for analyzing phylogenetic relationship among cultivated varieties, between tropical and temperate mulberry, for solving the vexed problem of identifying taxonomic positions of genotypes, for identifying markers associated with leaf yield attributing characters. As ISSR markers are one of the cheapest and easiest marker systems with high efficiency in generating polymorphism among closely related varieties, they would play a major role in mulberry genome analysis in the future.

Laser Captured Microdissection

  • 이경아
    • 한국동물학회:뉴스레터
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    • 제18권2호
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    • pp.21-25
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    • 2001
  • 대부분의 조직은 여러 가지 세포가 모여서 이루어지기 때문에 그 중의 어떤 특정세포에서 발현하는 물질을 분석하려면 조직을 이루고 있는 각각의 세포를 분리해내야 한다. 이렇게 순수하게 세포를 분리해내는 기술 중의 하나가 Laser Captured Microdissection (LCM)이 다. LCM의 개발로 기존에 사용되던 방법에 비하여 빠르고 간편하면서, 매우 정확하게 원하는 세포를 순수 분리해서 그 세포의 분자생물학적 또는 생화학적인 분석을 할 수 있게 되었다. LCM은 현미경으로 조직절편을 관찰하면서 원하는 세포를 낮은 에너지의 laser를 사용하여 도려내는 방법으로 조직절편 이외에도 도말된 혈액이나 자궁경부 조직, 그리고 배양된 세포를 cytocentrifugation한 후에 원하는 세포를 포획 할 수도 있다. LCM을 이용한 연구는 여러 분야에서 다양하게 진행되고 있으며, 특히 같은 조직 내에 존재하는 정상세포와 전이중인 세포, 그리고 암세포를 구분해 냄으로써 암의 전이기전 및 병인 연구에 매우 큰 공헌을 하고 있다. 이렇게 분리된 세포는 RT-PCR, LOH (loss of heterozygosity), microsatellite instability, differential gene profiling, cDNA microarray, Western blot, 2D PAGE protein analysis 등의 기법을 접목하여 연구하게 된다. 본 논단을 통하여 1996년 개발된 LCM의 원리와 이제까지 LCM을 이용한 연구 성과를 살펴보고자 한다.

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Genetic Diversity and Population Structure of Korean Soybean Collection Using 75 Microsatellite Markers

  • Lee, Gi-An;Choi, Yu-Mi;Yi, Jung-Yoon;Chung, Jong-Wook;Lee, Myung-Chul;Ma, Kyung-Ho;Lee, Sok-Young;Cho, Jin-Woong;Lee, Jung-Ro
    • 한국작물학회지
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    • 제59권4호
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    • pp.492-497
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    • 2014
  • Soybean (Glycine max L.) is crucial legume crop as source of high quality vegetable protein and oil, and Korea is regarded as a part of center of soybean origin. To expand the information of conserved genetic diversity, we analyzed the genetic variability of soybean collection mainly introduced Korean accessions using 75 microsatellite markers. A total of 1,503 alleles with an average value of 20.0 alleles were detected among 644 accessions. Korean collection revealed average allele number of 13.4 while Chinese, Japanese and Southeast Asian accessions showed 9.0, 5.4 and 6.5 mean alleles, respectively. Especially, Korean accessions showed more number of private allele per locus as 3.4 contrary to other geographical groups. The mean expected heterozygosity and polymorphic information content was 0.654 and 0.616, respectively, and expected heterozygosity values were not significantly distinguished according to the geographical groups. The phylogenetic dendrogram and deduced population structure based on DNA profiles of 75 SSR loci showed Korean accessions formed distinct gene pool against Chinese accessions, and could be divided into five subpopulations. Korean soybean accessions have specific genetic diversity and might be serve the valuable alleles for bio-industry as a part of the center of soybean origin.

Loss of ARID1A Expression in Gastric Cancer: Correlation with Mismatch Repair Deficiency and Clinicopathologic Features

  • Kim, Kyung-Ju;Jung, Hae Yoen;Oh, Mee-Hye;Cho, Hyundeuk;Lee, Ji-Hye;Lee, Hyun Ju;Jang, Si-Hyong;Lee, Moon Soo
    • Journal of Gastric Cancer
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    • 제15권3호
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    • pp.201-208
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    • 2015
  • Purpose: The AT-rich interactive domain 1A (ARID1A ) gene encodes BRG1-associated factor 250a, a component of the SWItch/Sucrose NonFermentable chromatin remodeling complex, which is considered a tumor suppressor in many tumors. We aimed to investigate the prognostic significance of ARID1A expression in gastric cancers and explore its relationship with clinicopathologic parameters such as mismatch repair protein expression. Materials and Methods: Four tissue microarrays were constructed from 191 resected specimens obtained at Soonchunhyang University Cheonan Hospital from 2006 to 2008. Nuclear expression of ARID1A was semiquantitatively assessed and binarized into retained and lost expression. Results: Loss of ARID1A expression was observed in 62 cases (32.5%). This was associated with more frequent vascular invasion (P=0.019) and location in the upper third of the stomach (P=0.001), and trended toward more poorly differentiated subtypes (P=0.054). ARID1A loss was significantly associated with the mismatch repair-deficient phenotype (P=0.003). ARID1A loss showed a statistically significant correlation with loss of MLH1 (P=0.001) but not MSH2 expression (P=1.000). Kaplan-Meier survival analysis showed no statistically significant difference in overall survival; however, patients with retained ARID1A expression tended to have better overall survival than those with loss of ARID1A expression (P=0.053). In both mismatch repair-deficient and mismatch repair-proficient groups, survival analysis showed no differences related to ARID1A expression status. Conclusions: Our results demonstrated that loss of ARID1A expression is closely associated with the mismatch repair-deficient phenotype, especially in sporadic microsatellite instability-high gastric cancers.

개에서 동경 수정란 이식 후 생산된 산자의 친자감별 (Parentage Testing for the Offspring Produced by Embryo Transfer with Frozen Embryo in the Dog)

  • 김용준;김하나;한용만;김선정;김병진;박영재;오홍근
    • 한국임상수의학회지
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    • 제17권1호
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    • pp.234-237
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    • 2000
  • The dornor, 2 years old, 20kg and mixed breed, was bred naturally on day 1 and day 3 of estrus and eight gastrulae were collected by flushing the uterus of the donor after laparatomy on day 13 after the second mating. The embryos were frozen by programmable freezer and preserved for about 3 months in liquid nitrogen. Another bitch in natural estrus, 2 years old, 30kg, mixed breed, was selected as the recipient and the frozen embryos(8 gastrulae) were thawed and each 4 embryos were transferred into upper partr of left and right uterine horn, respectively, on day 13 after the proper mating day determined by vaginal smear. The ecipient delivered 6 offspring 48 days after embryo transfer. Of 6 puppies, one was still birth and two puppies died one month after birth. Parentage test was performed by DNA analysis using microsatellite sequences for 3 puppiers, the recipient, the donor, the male dog bred with the donor, and the male dog raised near to the recipient. The markers selected for the test were CXX 873(133-157 base pair) and CXX 894(141-165 base pair). Using primers manufactured according to the markers, the blood samples were processed for polymerase chain reaction and the PCR products were treated for electrophoresis. The three puppies showed identical band to that of recipient, consequently, it was concluded that the puppies were offspring of the recipient mated naturally by the male dog, not the offspring by embryo transfer.

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No Genetic Differentiation of Elaphe schrenckii Subspecies in Korea Based on 9 Microsatellite Loci

  • An, Jung-Hwa;Park, Dae-Sik;Lee, Jung-Hyun;Kim, Kyung-Seok;Lee, Hang;Min, Mi-Sook
    • Animal Systematics, Evolution and Diversity
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    • 제26권1호
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    • pp.15-19
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    • 2010
  • The Russian ratsnake, Elaphe schrenckii, is found in Russia, China, and Korea, and is considered to be an endangered species by the Ministry of Environment in South Korea. Due to habitat loss and use in oriental medicine, their population has been severely decimated. In South Korea, two subspecies of E. schrenckii has been defined according to body color: E. s. schrenckii (blackish) and E. s. anomala (yellow-brownish). Molecular genetic studies on Elaphe schrenckii are very scarce and the taxonomy of Elaphe schrenckii subspecies is uncertain. From the present study, we attempted to identify the genetic differences of these two subspecies using species-specific microsatellites developed from the genomic library of E. schrenckii. Nine polymorphic loci were tested on 19 individuals from E. s. schrenckii (n=10) and E. s. anomala (n=9) in South Korea. The mean number of alleles was 3.78 in E. s. schrenckii and 4.11 in E. s. anomala. The average expected heterozygosity was 0.542 and 0.511 in E. s. schrenckii and E. s. anomala, respectively. We found a lack of genetic structure between two subspecies ($F_{ST}=0.016$) and no genetic discrimination between two subspecies was found. Based on the present findings by microsatellites, two subspecies can be considered as one species, E. schrenckii. However, further investigations on taxonomical status using mitochondrial and nuclear DNA sequences need to be performed and morphological & ecological data should be revised. The genetic markers should benefit future studies of the endangered species of other Elaphe species for the study of genetic diversity and potential conservation management.

Mitochondrial D-Loop Polymorphism and Microsatellite Instability in Prostate Cancer and Benign Hyperplasia Patients

  • Ashtiani, Zahra Ousati;Heidari, Mansour;Hasheminasab, Sayed-Mohammad;Ayati, Mohsen;Rakhshani, Naser
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.3863-3868
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    • 2012
  • In this study mitochondrial D-Loop variations in Iranian prostate cancer and benign prostatic hyperplasia (BPH) patients were investigated. Tumour samples and corresponding non-cancerous prostate tissue from 40 prostate cancer patients and 40 age-matched BPH patients were collected. The entire mtD-loop region (16024-576) was amplified using the PCR method and products were gel-purified and subjected to direct nucleotide sequencing. A total of 129 variations were found, the most frequent being 263A${\rightarrow}$G and 310T${\rightarrow}$C among both BPH and prostate cancer patients. Variation of 309 C${\rightarrow}$T was significantly more frequent in prostate cancer patients (P value<0.05). Four novel variations were observed on comparison with the MITOMAP database. Novel variations were np16154delT, np366G${\rightarrow}$A, np389G${\rightarrow}$A and 56insT. There was no correspondence between the different variations and the age of subjects. Considering that D-loop variations were frequent in both BPH and prostate cancer patients in our study, the fact that both groups had high average age can be a possible contributing factor. D-loop polymorphisms and microsatellite instability can influence cell physiology and result in a benign or malignant phenotype. Significantly higher frequency of 309 C${\rightarrow}$T variation in cancer patients is a notable finding and must be a focus of attention in future studies.