• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.897-902
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• 2004

To improve the methods used for the identification of Hanwoo, we performed a PCR using 3 -tailed primer associated with single nucleotide polymorphism(SNP) in Melanocortin 1 receptor(MCIR) gene. MCIR plays an important role in melanin synthesis and the SNP within MCIR was used as a target for PCRRFLP studies previously. A forward 3 -tailed primer, which matches with the template DNA of Hanwoo but not with others(blackhaired; Holstein and Black angus) at the site of 594th base sequence, and one reverse primer were designed for this study. When use this primer set, a size of 343bp was amplified by PCR only in Hanwoo, not in Holstein and Black angus. This result suggests that the PCR using our 3 -tailed primer would be very accurate, easy, reproducible and economic method to discriminate between Hanwoo meat and other black-haired ones.

• Journal of Life Science
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• v.24 no.8
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• pp.910-914
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• 2014

For the identification of the Jeju black cattle, Hanwoo and imported beef, we performed a multiplex polymerase chain reaction (PCR) associated with microsatellite (MS) and melanocortin 1 receptor (MC1R) gene. The MC1R gene plays an important role in regulation of the melanin synthesis within mammalian melanocytes. MC1R encoded by extension (E) locus was almost fixed with recessive red e allele in the Hanwoo. We estimated that the specific genotypes ($E^+/E^+$, $E^+/e$) of MC1R gene were characteristic genotypes of Jeju black cattle. But the PCR products resulted from using the MC1R gene derived primers only are not sufficient to identify Jeju black cattle from other relatives. We performed two times of successive multiplex PCR to provide a more accurate result for the identification of Jeju black cattle. The results suggest that two types of successive multiplex PCR methods using MC1R gene and Microsatellite derived primer set will be more useful to identification of Jeju black cattle, Hanwoo and imported beef.

• Korean Journal of Ecology and Environment
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• v.56 no.1
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• pp.1-13
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• 2023

Targeting Microcystin (MC), which is most abundantly detected in the North-Han River water area, we analyzed the relationship between the MC biosynthesis gene (mcyA gene), cyanobacteria cell density, and MC concentration, derived an RNA-MC conversion formula, and derived the cyanobacteria. The concentration of MC present in cells was predicted. In the North-Han River waters, the mcyA gene was found mainly at downstream sites of the North-Han River after Muk-Hyeon Stream junction, and higher copy numbers were found on average than other sites. In the Uiam Lake waters upstream of the North-Han River, the mcyA gene copy number increased at the Kong-Ji Stream point, and after September, the mcyA gene copy number decreased throughout the North-Han River waters. The expression of the mcyA gene was concentrated in the short period of summer due to the spatio-temporal difference between upstream and downstream water bodies. The mcyA gene expression level was not only highly correlated with MC concentration, but also correlated with the cell density of Microcystis aeruginosa and Dolichospermum circinale, which are known to biosynthesize MC. Six conversion formulas derived based on the RNA-MC relationship showed statistical significance (p<0.05) and exhibited high correlation coefficients (r) of 0.9 or higher. The expression level of MC biosynthesis gene present in eRNA determines the synthesis of cyanotoxin substances in water, quickly quantifies gene activity, and can be fully utilized for early warning of MC development.

• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.29-37
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• 2014

The Feather Color of chicken is considered as most obvious, and the purpose of this study is to identify the genotype following the SNP of MC1R, MITF and TYRP1, which are genes related to Feather Color, and develop a SNP marker that can be classified per breed. When a haplotype is observed through the combination of markers, a Korean Native Chicken can especially be distinguished when it is a CGG type in the SNP combination of the MC1R gene. In case of the TAG, TGG and TAA types, only Araucana was identified, and for the CAA type, Leghorn could specifically be distinguished. In the SNP combination of TYRP1 gene, only Leghorn was differentiated in case of the TTTCA and CCTCA types, and only Silky Fowl was identified in case of the CTTTA type. The SNP combination of MC1R gene enabled for Korean Native Chicken, Leghorn, and Araucana to be distinguished and each of the SNP and combination of TYRP1 gene allowed for all 4 breeds to be classified. If many researches are conducted about genetic polymorphism between breeds, then it is considered that the differences between breeds will be understood from a molecular biological aspect instead of simply distinguishing the breeds through Feather Color.

• Journal of the Korean Magnetic Resonance Society
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• v.16 no.1
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• pp.34-45
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• 2012

Melanocortin-4 receptor (MC4R) subtype is associated with obese humans. Especially, in a patient with severe early-onset obesity, novel heterozygous mutation in the MC4R gene was detected, resulting in an exchange of aspartic acid to asparagine in $90^{th}$ amino acid residue located in the predicted second trans-membrane domain (TM2). Mutations in the melanocortin-4 receptor (MC4R) gene are the most frequent monogenic causes of severe obesity which have been described as heterozygous with loss of function. In order to compare structure difference between MC4R wild type (MC4R-TM2-wt) and mutant (MC4R-TM2-D90N), we designed both MC4R-TM2-wt and MC4R-TM2-D90N construct in pET 21b vector. In this study, we optimized high-yield purification procedure for recombinant TM2-D90N. Eluted recombinant protein was resolubilized under urea condition for thrombin cleavage reaction and we conducted the high-performance liquid chromatography (HPLC) with reverse phase column under 1% acetonitrile, 0.01% TFA buffer solution. The molecular size of purified target peptide was confirmed by Tricine-SDS page analysis. To characterize MC4R-TM2-D90N, we have performed $^{15}N$-isotope labeling of peptide using M9 media and purified labeled target peptide for hetero-nuclear NMR spectroscopy.

• Korean Journal of Agricultural Science
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• v.38 no.3
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• pp.465-471
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• 2011

The commercial Korean native chickens (WR_CC) was developed by crossing a few native chicken breeds in Korea. In order to investigate the breed identification markers, SNPs from TYR gene and MC1R gene, which are associated with skin and feather colors respectively, were initially identified. In case of 3 identified SNPs in the TYR gene, yellow shank color was identified in Loss, Harvard, AA, RIR and CC, which have the fixed SNPs in most of the animals. On the other hand, SNP variations were observed in KNC_RB, C_B, WR_CC and HH_CC, which have the black, yellow and mixed color with black and yellow shank colors. Also, the investigation of 3 SNPs in the MC1R gene indicated that there were associations between shank and feather colors in RIR, SF, KNC_B, C_B and RIR. However, these results are not consistent among breeds. These SNP type inconsistencies within breeds suggested that the selection was performed based on the phenotypes, which is not include the genotype information. Thus, selection based on genetic information is required in the future.

• Journal of Animal Science and Technology
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• v.50 no.5
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• pp.633-640
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• 2008

With a double mismatch primer set designed for amplifying the modified DNA sequence fragments, bovine melanocortin-1 receptor(MC1R) gene encoded in Extension locus which plays a critical role in coat color development was analyzed using polymerase chain reaction mediated restriction fragment length polymorphism(PCR-RFLP). Amplified PCR fragments were successfully discriminated with combining the MspI- and AluI-RFLP into three major alleles(ED, E+, and e), directly related to bovine coat color phenotypes. The genotyping results showed that Jeju black cattle contained three MC1R alleles, but yellowish-red colored Hanwoo and bridle colored Korean Brindle cattle did not contained the dominant black allele ED. However, two dominant black-colored cattle breeds, Holstein and Angus, contained the ED allele over 96% in frequency. Hanwoo×Holstein F1 and Hanwoo×Angus F1 crossbred calves showed ED/e MC1R genotypes, and uniformly black coat color. the results suggested that this MC1R genotyping method be useful in allele discrimination for bovine MC1R gene which used for breed classification and characterization, as one of the important genetic markers, using combination of MspI- and AluI-RFLP for modified PCR product amplified with a newly designed double mismatch primer set.

• Journal of Animal Science and Technology
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• v.49 no.4
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• pp.437-442
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• 2007

The melanocortin-4 receptor(MC4R) is virtually expressed in all brain regions and plays an important role in energy homeostasis in mammals. MC4R has been intensively studied as a trait gene controlling economically important traits, such as growth and feed conversion, etc. Six hundreds and sixty Duroc pigs were genotyped for the MC4R locus and analyzed their relationships with breeding values for average daily gain(ADG), backfat thickness(BF), days to 90kg(D90) and feed conversion(FC). The estimated genotype frequencies for the all Duroc pigs were: 30.8%, 45.2%, 24.1% for AA, AB and BB genotypes, respectively. The mutant A allele was significantly associated with ADG, D90 and BF whereas no significant relationship was found with FC. The change of gene frequencies by generation was shown in both selected and culled groups. These results indicate that the MC4R polymorphism could be integrated in the present selection program to realize a marker-assisted selection for the growth traits of the Duroc population.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.5
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• pp.625-629
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• 2013

The melanocortin 1 receptor (MC1R) gene is related to the plumage color variations in chicken. Initially, the MC1R gene from 30 individuals was sequenced and nine polymorphisms were obtained. Of these, three and six single nucleotide polymorphisms (SNPs) were confirmed as synonymous and nonsynonymous mutations, respectively. Among these, three selected SNPs were genotyped using the restriction fragment length polymorphism (RFLP) method in 150 individuals from five chicken breeds, which identified the plumage color responding alleles. The neighbor-joining phylogenetic tree using MC1R gene sequences indicated three well-differentiated different plumage pigmentations (eumelanin, pheomelanin and albino). Also, the genotype analyses indicated that the TT, AA and GG genotypes corresponded to the eumelanin, pheomelanin and albino plumage pigmentations at nucleotide positions 69, 376 and 427, respectively. In contrast, high allele frequencies with T, A and G alleles corresponded to black, red/yellow and white plumage color in 69, 376 and 427 nucleotide positions, respectively. Also, amino acids changes at position Asn23Asn, Val126Ile and Thr143Ala were observed in melanin synthesis with identified possible alleles, respectively. In addition, high haplotype frequencies in TGA, CGG and CAA haplotypes were well discriminated based on the plumage pigmentation in chicken breeds. The results obtained in this study can be used for designing proper breeding and conservation strategies for the Korean native chicken breeds, as well as for the developing breed identification markers in chicken.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.68-69
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• 2005

To identify germline chimeric chicken using germ cell transplantation method, the testcross, spends much time, labor and cost to perform, is the only way for distinguishing germline chimeric chicken from normal one And to enhance the method, development of breed-specific molecular markers have been needed. We have just identified breed-specific sequence polymorphisms among Barred Plymouth rock, Korean Ogol Chicken and White Leghorn in PMEL17 and MC1R gene the loci of which are identical to dominant white and extended black loci. These sequence polymorphism will be very useful for screening germline chimera.