• Molecules and Cells
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• v.21 no.3
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• pp.411-417
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• 2006

We have developed a polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) marker that can distinguish male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker was located in a chloroplast psbA gene amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the restriction enzyme MspI revealed that N-cytoplasm plants have a functional MspI site (CCGG), whereas the S-cytoplasm plants has a substitution in that site (CTGG), and thus no MspI target. The results obtained using this PCR-RFLP marker to distinguish between cytoplasmic male sterile factors in 35 onion varieties corresponded with those using a CMS-specific sequence-characterized amplified region (SCAR) marker. Moreover, the PCR-RFLP marker can identify N- ot S-cytoplasms in DNA sample mixtures in which they are in up to a 10-fold minority, indicating that use of the marker has high diagnostic precision. We also demonstrated the usefulness of the SNP detected in the psbA gene for high-throughput discrimination of CMS factors using Real-time PCR and a TaqMan probe assay.

• Food Science of Animal Resources
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• v.25 no.2
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• pp.218-225
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• 2005

This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.

• Food Science of Animal Resources
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• v.24 no.4
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• pp.355-360
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• 2004

In Korean beef market, one of the major problems is mislabeling or fraudulent distribution of Holstein dairy meat or imported beef as domestic Hanwoo meat. Therefore, there has been a great need for a development of technology to identify beef breeds in meat and meat products. This study was carried out to develop the accurate and reliable method for the identification of beef breed using PCR-RFLP marker of MC1R, MGF and TYRPl genes affecting coat colors in cattle. A single base substitution (G\longrightarrowT transition) at the codon for amino acid position 104 of MC1R gene was identified between Hanwoo and Holstein and Angus breeds. The change at this position creates Msp I restriction site in Holstein and Angus, but not in Hanwoo. When the DNA amplified products (537 bp) was digested with Msp I, Hanwoo meat showed a single band of 537bp, while two fragments of 329bp and 208 bp were observed in Holstein meat and Angus breed, respectively. Thus, breed-specific RFLP marker in the MC1R gene can be used to distinguish between Hanwoo meat and Holstein and Angus meats. In the RFLP genotype of MGF gene, the frequency of r/r type was 75％ in Manwoo, whereas the frequency of R/R was 80％ in Hereford breed. Holstein and Angus breeds showed 100％ for R/r type. Therefore, Hanwoo meat showed significant difference in the MGF genotype frequencies compared with those of Holstein meat and imported beef cattle breeds. However, TYRP1 gene showed the same genotype in all breeds examined. Thus, this TYRP1 gene can not be used as a molecular marker for breed identification. As a consequence, we suggest that RFLP markers of the MC1R and MGF coat color genes could be used as DNA marker for identification of Hanwoo meat from Holstein and imported meats.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.05a
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• pp.195-198
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• 2005

본 연구는 cytochrome B 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 7종류 축종(닭, 양, 돼지, 소, 사슴, 말, 염소)의 육류로부터 cytochrome B 유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 축종의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(359 bp)을 두 종류의 제한효소(Hae Ш 및 Hinf I)로 각각 절단한 결과 축종 간 그리고 제한효소 간에 명확한 차이를 보이는 종 특이적인 PCR-RFLP marker를 검출하였다. 따라서, 본 연구에서 개발한 cytochrome B 유전자의 종 특이적 PCR-RFLP marker는 각종 식육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 DNA marker로 이용될 수 있을 것이다.

• Horticultural Science & Technology
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• v.30 no.1
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• pp.56-63
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• 2012

Tomato hypocotyl can generally be one of two colors, purple or green. Genetically, this trait is controlled by a single dominant gene. Hypocotyl tissue specific color expression is one of many visible genetic marker sources used to select tomato progeny. However, the visible marker does not show a clear distinction between homozygous genotype and heterozygous genotype from the breeding lines. Therefore, to identify a hypocotyl pigmentation related marker, we screened DNA polymorphisms in thirteen tomato lines showing purple or green hypocotyls. The markers used for screening consisted of primer set information obtained from anthocyanin related genes, conserved ortholog set II (COS II) marker sets localized near anthocyanin related genes, and restriction fragment length polymorphism (RFLP) markers localized near COS II markers, which produce polymorphisms between purple and green tomatoes. One primer from a RFLP fragment resulted in a polymorphism on agarose gel electrophoresis. From the RFLP fragment, a cleaved amplified polymorphic sequence (CAPS) marker was developed to distinguish between purple and green hypocotyls. The genotypes of 135 $F_2$ individuals were analyzed using the CAPS marker, and among them, 132 individuals corresponded to the phenotypes of hypocotyl pigmentation.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.05a
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• pp.199-202
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• 2005

본 연구는 cytochrome B 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 가금류 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 종간 근연관계가 높은 3종의 가금육(닭, 칠면조 및 오리)의 육류에서 분리한 DNA 시료를 대상으로 cytochrome B 유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 축종의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(359 bp)을 두 종류의 제한효소(HaeШ 및 Hinf I)로 각각 절단한 결과 특히, HaeШ 제한효소에서 가금류의 축종 간 그리고 제한효소 간에 명확한 차이를 보이는 종 특이적인 PCR-RFLP marker를 검출하였다. 따라서, 본 연구에서 개발한 cytochrome B 유전자의 가금류 종 특이적 PCR-RFLP marker는 가금류의 식육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 DNA marker로 이용될 수 있을 것이다.

• Journal of Life Science
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• v.13 no.3
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• pp.280-290
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• 2003

The incorporation of RAPD markers into the previous classical and RFLP genetic linkage maps will facilitate the generation of a detailed genetic map by compensating for the lack of one type of marker in the region of interest. The objective of this paper was to present features we observed when we associated RAPD map from an intraspecific cross of a Glycine max$\times$G. max, 'Essex'$\times$PI 437654 with the public RFLP map developed from an interspecific cross of G. max$\times$G. soja. Among 27 linkage groups of RAPD map, eight linkage groups contained probe/enzyme combination RFLP markers, which allowed us the incorporation of RAPD markers into the public RFLP map. Map position rearrangement was observed. In incorporating L.G.C-3 into the public RFLP linkage group a1 and a2, both pSAC3 and pA136 region, and pA170/EcoRV and pB170/HindIII region were in opposite order, respectively. And, pk400 was localized 1.8 cM from pA96-1 and 8.4 cM from pB172 in the public RFLP map, but was localized 9.9 cM from i locus and 18.9 cM from pA85 in our study. A noticeable expansion of the map distances in the intraspecific cross of Essex and PI 437654 was also observed. Map distance between probes pA890 and pK493 in L.G.C-1 was 48.6 cM, but it was only 13.3 cM in the public RFLP map. The distances from the probe pB32-2 to pA670 and from pA670 to pA668 in L.G. C-2 were 50.9 cM and 31.7 cM, but they were 35.9 cM and 13.5 cM in the public RFLP map. The detection of duplicate loci from the same probe that were mapped on the same or/and different linkage group was another feature we observed.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.34-34
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• 2003

한국산 선발계통 및 일본산 양식계통과 이들 두 계통간 잡종 참돔 집단의 유전적 특성을 분석하기 위하여, RAPD (Random Amplified Polymorphic DNA) marker를 탐색하였다. 10개의 염기로 이루어진 200개의 random primer 분석을 통하여 polymorphic pattern을 나타내는 23개의 random primer를 선발하였으며, 각 primer의 재현성을 확인하였다. 이들 중 OPA-11 primer는 크기가 각각 600 bp, 650 bp 및 750 bp 인 3개의 DNA 단편에 의하여 4개의 genotype을 나타냈으며, 각 genotype의 빈도는 집단간차이를 보였고, 한국산 선발계통 집단에서는 4개의 genotype이 모두 발견되는 반면, 일본산 양식계통 및 일본산 양식계통을 포함한 교배집단에서는 특정 genotype만 발견되었다. OPA-11 primer 유래의 polymorphic DNA 단편을 cloning하고 염기서열을 결정하였으며, SCAR (Sequence Characterized Amplified Region) primer를 제작하고 분석하였다. 본 연구는 참돔집단의 유전적 특성 파악 및 집단 구별에 RAPD marker를 활용하였으며, 참돔 육종시 형질 및 기능관련 DNA marker 탐색에 적용하기 위하여, 이후의 연구에서는 SCAR과 RFLP 분석에 RAPD marker를 이용하여 100% 정확도를 갖는 RFLP maker를 찾고, MAS (Marker-Assisted Selection)에 적용하고자 한다.

• Fisheries and Aquatic Sciences
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• v.9 no.3
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• pp.101-106
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• 2006

The cytochrome c oxidase subunit I (COI) gene region of mitochondrial DNA (mtDNA) was examined in four abalone species to estimate its utility as a genetic marker using restriction fragment length polymorphism (RFLP) analysis. The utility was evaluated in terms of genetic divergence and relationships among Haliotis discus hannai, H. rufescens, H. rubra, and H. midae in both hemispheres of the world. There was clear genetic divergence in the mtDNA COI region between all pairs of the four species. Moreover, relationships among the abalone species were reflected in their geographical distributions and morphological characteristics. Therefore, RFLP analysis of the mtDNA COI region is a suitable genetic marker for the estimation of genetic divergence and relationships among abalone species. However, it is not effective for the evaluation of genetic differences within abalone species.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2002.10a
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• pp.165-165
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• 2002

Hypertension is considered to be caused by a complicated combination of genetic and environmental factors. Atrial natriuretic peptide (ANP) has been to suppress renin activity and inhibit the synthesis and release of aldosterone. Therefore, Abnormalities of this peptide caused by genetic variation may be influence the blood pressure. The aim of present study was to examine the relationship between hypertension and Sca I RFLP of ANP gene in Korean population. The genotype distribution of this RFLP was significantly different between normotensives and hypertensives (P<0.05). However, this genetic marker was not significantly associated with any anthropometric parameters or plasma lipid concentrations in our study group. Therefore, our result suggest that Sca I RFLP of ANP gene may be useful as genetic marker in the ethiology of hypertension in Korean population, independent of any cardiovascular risk. factors studied.