• Title/Summary/Keyword: RFLP marker

검색결과 135건 처리시간 0.019초

Molecular Mapping of Resistant Genes to Brown Planthopper, Bphl and bph2, in Rice

  • Cha, Young-Soon;Cho, Yong-Gu;Shin, Kyeong-Og;Yeo, Un-Sang;Choi, Jae-Eul;Eun, Moo-Young
    • 한국작물학회지
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    • 제44권4호
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    • pp.345-349
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    • 1999
  • This study was carried out to map Bphl and bph2 gene in Mudgo and Sangju13 (Oryza sativa L.) respectively conferring resistance to brown plan-thopper (BPH) and to establish the marker-assisted selection (MAS) system. Bulked seedling (grown for 20 days) test was conducted with the 73 F4 lines derived from a cross between Nagdongbyeo and Mudgo for Bphl and with 53 BC3F5 lines derived from the Milyang95/Sangju13 cross for bph2. Bph1 was mapped between RG413 and RG901 on chromo-some 12 at a distance of 7.5 cM from RG413 and 8.4 cM from RG90l. A recessive gene bph2 was located near RZ76 on chromosome 12 at a distance of 14.4 cM. Bphl and bph2 were linked to each other with a distance of about 30 cM. An RFLP marker, RG413 linked to Bphl, was converted to an STS marker to facilitate the marker-assisted selection. BPH resistant genotypes could be selected with 92% accuracy in a population derived from a line of NM47-B-B.

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Identification of Korean Native Pork Using Breed-Specific DNA Marker of KIT Gene

  • Chung, Eui-Ryong;Chung, Ku-Young
    • 한국축산식품학회지
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    • 제30권3호
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    • pp.403-409
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    • 2010
  • Accurate methods for the identification of closely related species or breeds in raw and processed meats must be developed in order to protect both consumers and producers from mislabeling and fraud. This paper describes the development of DNA markers for the discrimination and improvement of Korean native pig (KNP) meat. The KIT gene is related to pig coat color and is often used as a candidate marker. A 538 bp fragment comprising intron 19 of the pig KIT gene was amplified by PCR using specific primers, after which the PCR amplicons of a number of meat samples from KNP and three major improved breeds (Landrace, Duroc and Yorkshire) were sequenced in order to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of two nucleotide substitutions, g.276G>A and g.295A>C, between KNP and the improved pig breeds. Digestion of KIT amplicons with AccII enzyme generated characteristic PCR-RFLP profiles that allowed discrimination between meats from KNP and improved pig. KNP showed three visible DNA bands of 264/249, 199, and 75 bp, whereas DNA bands of 249, 199, and 90 bp were detected in the three improved pig breeds. Therefore, the 75 bp DNA fragment was specific only to KNP, whereas the 90 bp DNA fragment was specific to the improved breeds. The breed-specific DNA markers reported here that target the KIT gene could be useful for the identification of KNP meat from improved pig meats, thus contributing to the prevention of falsified breed labeling.

Species Identification of Five Penaeid Shrimps Using PCR-RFLP and SSCP Analyses of 16S Ribosomal DNA

  • Khamnamtong, Bavornlak;Klinbunga, Sirawut;Menasveta, Piamsak
    • BMB Reports
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    • 제38권4호
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    • pp.491-499
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    • 2005
  • DNA-based molecular markers for differentiation of five penaeid shrimps (Penaeus monodon, P. semisulcatus, Feneropenaeus merguiensis, Litopenaeus vannamei and Marsupenaeus japonicus) were developed based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-stranded conformation polymorphism (SSCP) of 16S ribosomal (r) DNA. Differentiation of P. monodon, P. semisulcatus and L. vannamei can be unambiguously carried out by PCR-RFLP of 16S $rDNA_{560}$ whereas P. semisulcatus and M. japonicus shared a BABB mitotype. These shrimps were successfully discriminated by SSCP analysis of 16S $rDNA_{560}$. Nevertheless, the amplification success for L. vannamei and F. merguiensis was not consistent when tested against larger sample sizes. As a result, 16S $rDNA_{560}$ of an individual representing the most common mitotype of each species was cloned and sequenced. The new primer pair was designed and tested against the large sample sizes (312 bp product, N = 185). The amplification success was consistent across all species. PCR-RFLP of 16S $rDNA_{312}$ was as effective as that of 16S $rDNA_{560}$. Differentiation of all shrimp species were successfully carried out by SSCP analysis.

콩 Cyst 선충 Race 5에 대한 저항성 QTL 탐색 (Identification of Quantitative Trait Loci for Resistance to Soybean Cyst Nematode Race 5)

  • Choi, In-Soo;Kim, Yong-Chul;Kim, Sung-Man;Lee, Chung-Yeol;Park, Hyean-Cheal;Halina T. Skorupska
    • 한국작물학회지
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    • 제42권6호
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    • pp.712-721
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    • 1997
  • 콩 품종 Essex와 PI 437654간 교잡 후 F$_2$유래 F$_3$ 계통들을 재료로 하여 작성된 RAPD 유전자지도상에 cyst 선충 race 5에 대한 저항성 QTLs 분석을 실시한 바 결과를 요약하면 다음과 같다. 1. 회귀분석 결과 26개의 marker들(22 RAPD, 4 RFLP)에서 cyst 선충 race 5 저항성 반응에 대한 유의성이 인정되었다. 2. MAPMAKER /QTL 분석 결과2개의 저항성 QTL들이 탐색되었는데, 이 QTL들은 2개의 linkage groups(LGC-20와 Group 2)에 위치하였다. 3. 탐색된 2개의 QTL들 중 1개는 우성유전, ?고 나머지 하나는 열성유전양상을 나타내었다. 4. 콩 cyst 선충 race 5의 저항성에 대한 유의성이 인정되는 5개의 marker들간 상호작용을 알아보기 위한 다중회귀분석 결과 총 26개의 조합들 중 4개의 marker들(E02$^3$, G$10^1$, W03, pK418C)로 구성된 조합에서 가장 높은 표리적 변이의 값(35.2%)을 나타내었다.

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DNA Analysis of mtDNA COI Gene in the Sharp-toothed Eel (Muraenesox cinereus Forskal) from Yeosu, Jinhae, Jeju, Goseoung, Jangheung and Haenam Populations in Korea Using PCR-aided RFLP

  • Oh, Taeg-Yun;Jeong, Sun-Beom;Cho, Eun-Seob
    • 한국환경과학회지
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    • 제20권4호
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    • pp.551-554
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    • 2011
  • The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

Characterization of MHC DRB3.2 Alleles of Crossbred Cattle by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

  • Paswan, Chandan;Bhushan, Bharat;Patra, B.N.;Kumar, Pushpendra;Sharma, Arjava;Dandapat, S.;Tomar, A.K.S.;Dutt, Triveni
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권9호
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    • pp.1226-1230
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    • 2005
  • The present investigation was undertaken to study the genetic polymorphism of the DRB3 exon 2 in 75 crossbred cattle by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Five genotypes i.e. HaeIII-a, HaeIII-b, HaeIII-e, HaeIII-ab and HaeIII-ae were observed when the 284 bp PCR products were digested with HaeIII restriction enzyme. The corresponding frequencies of these patterns were 0.53, 0.04, 0.01, 0.38 and 0.04, respectively. Digestion with RsaI restriction enzyme resolved 24 different restriction patterns. The frequencies of these patterns ranged from 0.013 (RsaI-f, RsaI-k and RsaI-c/n) to 0.120 (RsaI-n). The results revealed that the crossbred cows belonged to the RsaI patterns namely b, k, l, a/l, d/s, l/n, l/o and m/n, whose corresponding frequencies were 0.027, 0.013, 0.040, 0.027, 0.040, 0.067, 0.027 and 0.067, respectively. Digestion of the 284 bp PCR product of DRB3.2 gene with PstI in the crossbred cattle did not reveal any restriction site. These results suggested the absence of the recognition site in some of the animals. These results also revealed that the crossbred cows studied were in homozygous as well as heterozygous condition. On the basis of the above results it can be concluded that the DRB3.2 gene was found to be highly polymorphic in the crossbred cattle population.

벼 흰잎마름병 저항성 유전자(Xa-1, Xa-3)연관 RFLP 마커 탐색 (Mapping of RFLP Markers Linked to Bacterial Blight Resistant Genes (Xa-1, Xa-3) in Rice)

  • 강현중;김현순;남정권;이영태;이승엽;김석동
    • 한국작물학회지
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    • 제48권6호
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    • pp.419-423
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    • 2003
  • 우리나라 자포니카 품종의 흰잎마름병 저항성 유전자와 연관된 마커를 탐색하기 위하여, 밀양121호, 밀양123호 및 HB10624-AC5 등을 교배친으로 한 두 조합의 약배양 계통을 재료로 흰잎마름병 저항성 유전자(Xa-1 and Xa-3)와 DNA 마커간의 연관분석을 통하여 유전자 지도를 작성하고자 수행하였던 결과를 요약하면 다음과 같다. 1. $\textrm{K}_1$ 균주에 대한 흰잎마름병 저항성 검정결과, 밀양121호/HRl1650-1-4-2에서는 저항성과 감수성이 1:1로 분리하였으며, 밀양123호/HR10624-AC5 조합의 $\textrm{K}_1$$\textrm{K}_3$ 균주에 대한 검정결과는 각각 3:1과 1:1로 분리하여 이론치에 합당하였다. 2. 교배친에 대하여 DraI. HindIII, EcoRI, EcoRV, PstI등 5가지 제한효소에 대한 다형현상을 검정한 결과, RZ590, RG303, RZ536 등 3개의 마커가 다형현상을 나타내었다. 3. 흰잎마름병 포장저항성 검정결과와 RFLP 마커와의 연관분석 결과 Xa-1 유전자는 RZ590과 4번 염색체 상에서 3.1$\times$1.5 cM으로 연관되어 있었으며, Xa-3 유전자는 Rz536 및 RG303과 11번 염색체 상에서 각각 7.6$\times$2.3 및 16.0$\times$3.2 cM으로 연관되어 있었다. 4. 11번 염색체 상에서 Xa-3와 Rz536 및 RG303은 "Xa-3-RZ536-RG303" 순으로 위치하였다.순으로 위치하였다.

현삼속 식물의 종판별을 위한 Mitochondrial DNA의 염기서열 및 PCR-RFLP 분석 (Molecular Authentication of Scrophularia herbs by PCR-RFLP Based on rpl-5 Region of Mitochondrial DNA)

  • 이정훈;조익현;이제완;박춘근;방경환;김홍식;박충범
    • 한국약용작물학회지
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    • 제18권3호
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    • pp.173-179
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    • 2010
  • This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.