• Title/Summary/Keyword: 분자 성판별

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Identification of Raw Materials in Processed Meat Products by PCR Using Species-Specific Primer (종 특이 프라이머를 이용한 식육가공품의 사용원료 판별법)

  • Park, Yong-Chjun;Ahn, Chi-Young;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Park, Kun-Sang;Yoon, Hae-Sung
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.68-73
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    • 2012
  • In this study, a method was developed using molecular biological technique to distinguish an authenticity of meats for processed meat products. The genes for distinction of species about meats targeted at 12S or 16S genes in mitochondrial DNA and the species-specific primers were designed by that PCR products' size was around 200bp for applying to processed products. The target materials were 10 species of livestock products and it checked whether expected PCR products were created or not by electrophoresis after PCR using species-specific primers. The results of PCR for beef, pork, goat meat, mutton, venison, and horse meat were 131, 138, 168, 144, 191, and 142 bp each. The expected PCR products were confirmed at 281, 186, 174, and 238 bp for chicken, duck, turkeymeat, and ostrich. Also, non-specific PCR products were not detected in similar species by species-specific primers. The method using primers developed in this study confirm to be applicable for composite seasoning including beefs and processed meat products including pork and chicken. Therefore, this method may apply to distinguish an authenticity of meats for various processed products.

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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Analysis of Genetic Polymorphisms and Similarity Using Random Amplified Polymorphic DNAs in Cattle (RAPD기법을 이용한 축우의 유전적 다형성과 유사도 분석)

  • Lee, S.H.;Seo, K.W.;Kwon, I.;Sung, C.K.;Kim, S.K.;Sang, B.C.
    • Korean Journal of Agricultural Science
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    • v.26 no.2
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    • pp.39-48
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    • 1999
  • This study was carried out to investigate the difference and genetic similarity at the level of molecular genetics. Genomic DNA was extracted from blood of Holstein, Korean cattle, Charolais, and hybrid between Korean cattle and charolais and RAPD(random amplified polymorphic DNAs) was analyzed by PCR(polymerase chain reaction). After genetic similarity value from different breeds are analyzed, genetic similarity was estimated by UPGMA(unweighted pair-group method using average). The results obtained from this study can be summarized as follows: 1. When genomic DNA which was extracted from different breeds was subjected to electrophoresis on 1.5% agarose gel, bigger than 12.2kb was appeared. Ratio by absorbance of $A_{260}/A_{280}$ was 1.75~2.10, indicating that genomic DNA was quite pure for RAPD analysis. 2. Different band patterns by RAPD were appeared according to the breeds in cattle. The best primer used to distinguish Holstein from other breeds was 5'-GAC CGC TTG T-3'. 3. A 340bp fragment was amplified in $33.0^{\circ}C$ of annealing temperature for the Holstein and Charolais breeds, but any amplification was not occurred in this annealing temperature for Korean cattle and hybrid. In addition, a 340bp fragment was amplified in $37.5^{\circ}C$ of annealing temperature for the Holstein and Korean cattle, but any amplification was not occurred in this annealing temperature for Charolais and hybrid. For the reaction of PCR. $37.5^{\circ}C$ and $33.0^{\circ}C$ of annealing temperature was shown to be best for genetic marker identification from Holstein, Charolais, and hybrid between Korean cattle and Charolais. 4. When genetic similarity from different breeds are analyzed at the both temperature of $33.0^{\circ}C$ and $37.5^{\circ}C$, the genetic similarity value of Holstein and Korean cattle, Holstein and Charolais, Korean cattle and Charolais, and Korean cattle and hybrid were 0.666~0.777, 0.615~0.666, 0.400~0.461 and 0.857~0.888, respectively. 5. It could be concluded that different breeds are capable of distinguishing by RAPD used random primer 5'-GAC CGC TTG T-3', genetic similarity from different breeds was appeared the higher genetic similarity value of Korean cattle and Charolais than that of Holstein between Korean cattle and Charolais by UPGMA.

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Species-specific Marker of Phytophthora pinifolia for Plant Quarantine in Korea (국내 식물검역대상 Phytophthora pinifolia의 PCR 검출을 위한 종 특이적 마커 개발)

  • Kim, Narae;Choi, You Ri;Seo, Mun Won;Song, Jeong Young;Kim, Hong Gi
    • The Korean Journal of Mycology
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    • v.44 no.2
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    • pp.103-107
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    • 2016
  • To establish a rapid and accurate detection of Phytophthora pinifolia, which is a quarantine pathogenic fungus in Korea, a species-specific primer was developed based on the ras-related protein (Ypt1) gene. Species-specific primer based on the DNA sequences of Ypt1 gene amplified 193 bp polymerase chain reaction (PCR) product for P. pinifolia. The primer pair yielded the predicted PCR product size exactly in testing with target pathogen DNAs, but not from the other 10 species of Phytophthora and 14 species of other phytopathogenic fungi. The primer pair also showed only the species-specific amplification curve on realtime PCR on target pathogen DNA. The detection sensitivity of real time PCR using species-specific primer pair was 10 to 100 times higher than conventional PCR, with 1 to $10pg/{\mu}L$.

Genetic analysis of endangered species Crested Ibis (Nipponia nippon) microsatellite markers (Microsatellite Markers를 이용한 따오기의 유전적 특성 분석)

  • Kim, Da Hye;Kim, Yi Seul;Seo, Joo Hee;Kim, Sung Jin;Kong, Hong Sik
    • Korean Journal of Ornithology
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    • v.25 no.2
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    • pp.77-81
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    • 2018
  • The Korean Crested ibis Nipponia Nippon is an endangered species. A pair of Crested ibis was introduced from China in October 2008, and a successful program of artificial incubation of the species, and over 200 animals have been successfully bred through the restoration project up to 2017 at Upo ibis restoration center. We assessed genetic diversity and sex determination in the Korean Crested ibis. In total, 228 Crested ibis (115 females and 113 males) were identified. And genetic diversity measures, observed heterozygosity, expected heterozygosity, and polymorphic information content values were lower in 2017 than those in 2016. The inbreeding coefficient showed that the degree of ancestry increased in 2017. The decrease in polymorphism and increase in the degree of ancestry is thought to be due to inbreeding in such a small group. In this study provided important insight into protocols for genetic management of the breeding population of Korean Crested ibis in Korea and will help in extending the restoration program.

GC-MASS Analysis and Microbial Enumeration for the Identification of Spoiled Red Pepper Powder (GC-MASS 분석과 미생물 균수 차이에 의한 희아리 고춧가루 판별)

  • Jeong, Su-Jin;Han, Sang-Bae;Uhm, Tai-Boong
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.191-197
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    • 2008
  • This work was intended for the identification of markers that are found only in the spoiled red pepper powder. When analyzed by GC/MASS, the spoiled red pepper powder contains characteristic naphthalene derivatives, 1, 2, 3, 5, 6, 7, 8, $8\alpha$-octahydro-1, $8\alpha$-dimethyl-7-(1-methylethenyl)-naphthalene and 2-isopropenyl-$4\alpha$, 8-dimethyl-1, 2, 3, 4, $4\alpha$, 5, 6, $8\alpha$-octahydronaphthalene, which have not found in the normal red pepper powder. In addition, microscopic observation and microbial enumeration of the red pepper powder had been performed. Images by scanning electron microscopy showed that the surfaces of spoiled pepper powder were rough with many kinds of microbes, compared with those of normal red pepper powder. A good correlation between the bacterial and fungal counts in the same sample was observed and could be clearly classified into two groups, the normal and the spoiled group, by difference in the microbial counts. These results suggest that the spoiled red pepper powder can be identified by a combination of GC/MASS, microbial counts, and scanning electron microscopy.

Identification of new Breeding Lines by Prunus Persica Cultivar-Specific SCAR Primers (SCAR 마커 개발 및 이를 활용한 국내 육성 복숭아 품종 판별)

  • Han, Sang Eun;Cho, Kang-Hee;Nam, Eun Young;Shin, Il-Sheob;Kim, Chung Hee;Kim, Hyun Ran;Kim, Dae-Hyun
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.495-501
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    • 2010
  • Peaches (Prunus persica) are less popular than the fresh fruits, because their flesh gets soft faster. So many breeders focused on their aim to firmness. Other breeders focused on juiciness, flavor and aroma. Breeding requires much labor, time and money. To reduce these requirements, many scientists develop many SSR, CAPS and SCAR makers. New peach varieties bred in our National Institute of Horticultural & Herbal Science (NIHHS) such as, Cheonhong, Suhong and Harhong are yellow flesh cultivars and Yumyeong, Baekmijosaeng, Baekhyang, Jinmi, Soomee, Mihong, Misshong and Yumee are white flesh cultivars. These peach cultivars are planted in orchard of Korea. To assert breeding cultivar patents and prevent patent disputes, we detected cultivar-specific DNA fragment using 235 sets of Operon RAPD primers, analyzed 134 DNA sequences and constructed SCAR primers. To confirm the cultivar-specific SCAR markers, we applied candidate SCAR primers to 30 peach cultivars widely cultivated in Korea. These selected lines are included father and mother lines that were used to develop new varieties in NIHHS. Using fourteen SCAR primer sets, we characterized thirty cultivars selected. The SCAR marker is expected to serve as molecular evidence distinguishing different peach varieties.

Population Structure of Korean Paraplagusia japonica (Cynoglossidae) Based on Morphological and Molecular Markers (한국산 흑대기 Paraplagusia japonica (참서대과)의 형태 및 분자 마커에 의한 집단구조)

  • Park, Gyeong Hyun;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.34 no.2
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    • pp.73-85
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    • 2022
  • The cynoglossid fishes are popular for food in the world including Korea, China and Japan, and among them, Paraplagusia japonica lives all over the sea of Korea. In order to establish appropriate management measure, it is essential to clarify population structure of P. japonica from the morphological and molecular perspectives. We collected a total of 132 individuals of P. japonica from six localities in Korea between 2008 and 2021. Canonical discriminant analysis results showed that the West Sea population (Incheon) slightly differed from the South (Tongyeong, Busan) and East Sea populations (Pohang, Donghae, Sokcho). Similar results were also shown in Kruskal-Wallis test of meristic characters. Furthermore, neighbor-joining and maximum-likelihood trees based on 849 base pairs of mitochondrial DNA cytochrome b sequences showed that P. japonica was divided into two lineages (designated as A and B) with a high significance (Φst=0.0781, P<0.001). Interestingly, however, the two lineages in the admixture area (South-East Sea) were not different in morphological characters. Our results suggest that P. japonica had undergone differentiated history during the Late Pleistocene, but secondary contact may occur at the admixture area.

Improvement of Seedling Establishment in Wet Direct Seeding of Rice using the Anaerobic Germination Tolerance Gene Derived from Weedy Photoblastic Rice (잡초벼 PBR 혐기발아 내성 유전자 활용 벼 담수직파 초기 입모 개선)

  • Jeong, Jong-Min;Mo, Youngjun;Baek, Man-Kee;Kim, Woo-Jae;Cho, Young-Chan;Ha, Su-Kyung;Kim, Jinhee;Jeung, Ji-Ung;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.161-171
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    • 2020
  • Direct seeding is one of the rice seedling establishment methods that is increasingly being practiced by farmers to save labor and reduce costs. However, this method often causes poor germination under flooding conditions after sowing. In this study, we developed japonica elite lines with quantitative trait loci (QTL) associated with anaerobic germination (AG) tolerance to overcome poor germination and seedling establishment in wet direct seeding. The QTL introgression lines were developed from a cross between weedy photoblastic rice as the AG donor and the Nampyeong variety via phenotypic and genotypic selection. Compared to Nampyeong, the survival rates of the selected lines were improved by approximately 50% and 240% under field and greenhouse conditions, respectively. To improve selection efficiency by marker assisted selection, the QTL markers associated with AG tolerance were converted to cleaved amplified polymorphic sequence markers designed based on next-generation sequence analysis. These lines retained similar agronomic traits and yield potential to the parent, Nampyeong. Among these lines, we selected the most promising line, which exhibited high survival rate and good agricultural traits under flooding conditions and named the line as Jeonju643. This line will contribute to breeding programs aiming to develop rice cultivars adapted to wet direct seeding. This study demonstrates the successful application of marker-assisted selection to targeted introgression of anaerobic genes into a premium quality japonica rice variety.

Discrimination and Hordein Polypeptide Patterns of Malting Barley Varieties Using UPLC (UPLC 분석을 이용한 맥주보리 품종의 호데인 단백질 분석 및 품종 판별)

  • Yoon, Young-Mi;Kim, Yang-Kil;Kang, Chon-Sik;Park, Jin-Cheon;Park, Tae-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.326-338
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    • 2021
  • Ultra-performance liquid chromatography (UPLC) was used to assess the hordein protein fraction of malting barley. C-hordeins (barley prolamins) were extracted with 70% ethanol (EtOH) and 55% isopropyl alcohol (IPA, 2-propanol), and B-hordeins were extracted with the same alcohols in 1.0% dithiothreitol (DTT). High molecular weight (HMW) prolamins (D-hordeins) were extracted with 50% IPA with 1M Tris-HCl (pH 8.0). The same protein patterns were observed in both the experimental extraction solutions (EtOH and IPA). However, the patterns of hordein, extracted with EtOH and IPA containing 1.0% DTT, differed slightly. C- and B-hordeins extracted from those solutions were analyzed. Twenty-six malting barley varieties developed in Korea were analyzed using UPLC. The varieties were divided into seven groups according to hordein patterns of retention time 16 min to 18 min, and 20 varieties showed unique patterns.