한국재래산양 β-lactoglobulin 유전자 5'flanking 영역의 염기서열 분석

류승희,한성욱,서길웅,상병찬 충남대학교 농업과학연구소 2001 농업과학연구 Vol.28 No.2

본 연구는 한국 재래산양의 β-lactoglobulin(β- LG) 유전자 발현조절부위의 특성을 구명하기 위하여 PCR기법으로 specific primer를 이용하여 β-LG 발현조절부위를 증폭한 후 염기서열을 분석하여 Sheep종의 β-LG유전자 발현조절부위의 염기서열상의 차이를 분석하였다. 한국 재래산양의 genomic DNA로부터 PCR기법을 이용하여 β-LG 유전자 발현조절 부위를 증폭한 결과 Sheep종에서 보고된 바와 같이 1,077bp의 단편크기로 증폭되었음을 확인 할 수 있었다, Sheep종에서 보고된 β-LG 발현조절 부위의 염기서열과 한국재래산양에서 분석된 염기서열간의 차이는 총 897개의 염기중 46개의 nucleotide에서 차이를 나타내어 94.9%의 상동성을 보였다. 특히 한국재래산양과 Sheep종간 염기서열상의 차이는 Sheep종의 T염기가 재래산양의 C염기로 치환되었거나 반대로 Sheep종의 C염기가 재래산양에서 T염기로 치환되어 있음을 확인할 수 있었다. 이상의 결과에서 β-LG유전자 발현조절부위의 염기서열은 재래산양과 Sheep 종간에 높은 상동성을 보였으나 이들 종간의 발현조절부위의 염기서열상의 차이에 대한 유전자 발현조절 관계와 전사요소는 앞으로 연구가 더 진행되어야 할 것으로 사료된다. This study was analyzed by PCR technique with specific primer in order to investigate the characteristics of β-lactoglobulin (β-LG) gene 5'flanking region in Korean native goat. This work confirmed amplified product of 1,077 bp fragments obtained from the amplification of β-LG promoter from genomic DNA using PCR in Korean native goat. The nucleotide sequence of β-LG gene 5'flanking region in Korean native goat as compared with that of sheep β-LG were different at 46 base of 897 nucleotides, and showed high homology as about 94.9% each breed. Especially we confirmed that the difference of nucleotide sequences between Korean native goat and sheep were consisted of T→C substitution and C→T substitution reversely. As a consequences, the sequences of β-LG gene 5'flanking region showed a high homology between Korean native goat and sheep. Furthermore we should be studied that relationships between the control of gene expression and nucleotide sequences of transcription factor in Korean native goat.

PCR 기법을 이용한 한국재래산양 β-casein 유전자의 특성

김지애,류승희,유성란,이준헌,서길웅,김선균,상병찬 충남대학교 농업과학연구소 2002 농업과학연구 Vol.29 No.2

본 연구는 한국 재래 산양 112두와 유산양인 Saanen종 7두의 혈액으로부터 genomic DNA를 추출하고, PCR-RFLP 방법에 의해 β-casein 유전자의 특성을 분석하여 한국재래산양의 효율적인 유전자원의 보전 및 개량을 위한 기초 자료로 제공하고자 실시하였다. 한국재래산양의 genomic DNA로부터 PCR기법을 이용하여 β-casein의 유전자좌를 증폭한 결과 각각 481bp 크기의 단편이 양호하게 증폭되었음을 확인하였다. β-casein 유전자좌의 증폭산물에 대한 Bal Ⅰ의 제한효소를 처리한 결과, β-casein AB형은 481bp, 284bp 및 197bp의 단편을, 그리고 BB형은 284BP와 197BP의 단편을 한국재래산양과 유산양인 Saanen 종에서 확인 할 수 있었다, 유전자형 빈도에 있어서는 한국재래산양에서 β-casein AB 및 BB의 빈도는 각각 6.25 및 93.75%이었고, 유산양인 Saanen 종은 각각 57.14 및 42.86%이었다. 유전자빈도에 있어서는 한국재래산양의 β-casein A 및 B의 빈도가 각각 0.031 및 0.969이었고, Saanen 종에서는 각각 0.286 및 0.714의 빈도를 보였다 한국재래산양의 β-casein 유전자의 염기서열과 이미 보고되어 있는 goat의 염기서열(GeneBank accession Number M90556)간에는 총 11개의 염기서열에 차이를 나타내어 97.71%의 상동성을 보였다, 따라서 한국재래산양의 β-casein 유전자의 다형성과 염기서열 분석에 의한 분자유전학적 특성의 규명은 한국재래산양의 유전자원의 보전 및 개량을 위한 기초 및 응용 자료로 이용될 수 있을 것으로 생각된다. This study was performed to provide the basic data for preservation and improvement of genetic resources according to finding genetic construction obtained from analysis of genetic characteristics of β-casein gene in Korean Native goat and Saanen using the PCR-RFLP. This study confirmed the amplified products of 481bp fragments obtained from the amplification of β-casein loci by PCR. The β-casein AB genotype showed 481, 284 and 197bp, and β-casein BB genotype showed 284 and 197bp fragments in Korean Native goat and Saanen. The frequencies of β-casein genotype in Korean Native goat were 6.25 and 93.75% for AA and AB and the frequencies of β-casein genotype in Saanen were 57.14 and 42.86% for AA and AB types. The frequencies of β-casein A and B alleles were 0.031 and 0.969 in Korean Native goat and the frequencies of β-casein A and B alleles are 0.286 and 0.714 in Saanen, respectively. The nucleotide sequence of β-casein gene of Korean Native goat was 97.71% higher homology with 11 nucleotide sequences difference of that of goat reported in GeneBank (M90556). Therefore, this study of molecular genetic characteristics by the analysis of genetic polymorphism and sequencing for β-casein gene should be used as basic and applying data for preservation and improvement of genetic resources in Korean Native goat breeding.

PCR 기법을 이용한 축우의 β-lactoglobulin 및 κ-casein 유전자형 분석에 관한 연구

상병찬,류승희,이상훈,송치은,남명수,전병순 충남대학교 농업과학연구소 1998 농업과학연구 Vol.25 No.2

This study was performed to offer the basic and applicable data for improvement of Korean cattle and dairy cattle, according to finding the genetic construction obtained from analysis of genetic polymorphisms of β-lactoglobulin and κ-casein loci related Korean cattle and Holstein cows using PCR-RFLP. Genomic DNA used in this study was prepared from the blood of 253 individuals of Korean cattle in Korean Native Cattle Improvement Center, NLCF, and the blood of 113 individuals of Holstein cows in National Livestock Research Institute. The results obtained are summarized as follows : 1. This study confirmed amplified products of 530bp and 262bp fragments obtained from the amplification of β-lactoglobulin and κ-casein loci in Korean cattle and Holstein breed by PCR. 2. The β-lactoglobulin AA genotype showed 153bp and 109bp fragments, and β-lactoglobulin AB genotype showed 153bp, 109bp, 79bp and 74bp fragments, and BB genotype showed 109bp, 79bp and 74bp fragments in amplified products of β-lactoglobulin loci with the restricted enzyme digestion of Hae III. 3. The κ-casein AA genotype showed a 530bp fragment, and κ-casein AB genotype showed 530bp, 344bp and 186bp fragments, and BB genotype showed 344bp and 186bp fragments in amplified products of κ-casein loci with the restricted enzyme digestion of Taa I. 4. On β-lactoglobulin genotypes and gene frequencies, Korean cattle were 6.72%, 26.09% and 67.19% for AA, AB and BB genotypes, and β-lactoglobulin A and B alleles were 0.197 and 0.803, and Holstein were 35.40%, 56.64% and 7.96% for AA, AB and BB genotypes, and β-lactoglobulin A and B alleles were 0.637 and 0.363, respectively. 5. On κ-casein genotypes and gene frequencies, Korean cattle were 46.25%, 39.13% and 14.62% for AA, AB and BB genotypes, and κ-casein A and B alleles were 0.658 and 0.342, and Holstein were 60.18% and 38.94% and 0.88% for AA, AB and BB genotypes, and κ-casein A and B alleles were 0.796 and 0.204, respectively. 6. As a consequence, the gene frequency was 0.197 and 0.803 for β-lactoglobulin A and B alleles, and 0.658 and 0.342 for κ-casein A and B alleles in Korea cattle, but was 0.637 and 0.363 for β-lactoglobulin A and B alleles, and 0.796 and 0.204 for κ-casein A and B alleles in Holstein, respectively.

한우와 Holstein종의 k -casein 유전자의 발현조절부위의 염기서열 분석

상병찬,류승희,이상훈 충남대학교 농업과학연구소 2000 농업과학연구 Vol.27 No.1

본 연구는 한우와 Holstein종의 κ-CN 유전자발현조절부위, 즉 κ-CN gene 5'-flanking region에 대한 특성을 구명하기 위하여 specific primer를 이용하여 PCR(polymerase chain reaction) 방법으로 분석하였다 PCR 증폭 결과, 349bp 크기의 증폭산물을 얻었으며 한우와 Holstein종의 π-CN 운전자의 발현조절부위의 염기서열 분석결과에 있어서는 -82bp에서 -431bp까지 분석되었으며, 종간의 염기서열 비교에서 -386bp에서는그olstein종에서는 T 염기가 한우에서는 C 염기로치환되었으며, -241bp와 -192bp에서는 Holstein종에서는 T 및 C 염기가 존재하였으나, 한우에서는 결실되었다. 또한, -183bp에서는 한우에서는 T염기가 존재하였으나, Holstein종에서는 염기가 결실되어 있음을 알 수 있었다 또한 한우와 Holstein종간의 상동성 분석에 있어서는 한우 κ-CN 유전자 발현조절부위의 염기서열이 Holstein종과98.9%의 상동성을 보였다. This study was carried out to investigate the characterizations of κ -CN genes of milk protein. In order to find out the characterizations of κ -CN genes, the nucleotide sequences of 5'-flanking regions of κ -CN genes were analyzed by PCR (polymerase chain reaction) technique with specific primers in order to investigate the characterizations of these promoters in Korean Native cattle and Holstein. PCR products obtained 349bp fragment and the nucleotide sequences of Korean Native cattle and Holstein of κ -CN gene 5'-flanking region was analyzed to -82bp from -431bp. On the comparison of each breed, Holstein substituted T→C at -386bp, and -241bp(T) and -192bp(C) existed, but Korean Native cattle was deleted. Also, Korean Native cattle was existed T at -183bp but Holstein was not. The homology analysis of between Korean Native cattle and Holstein was showed 98.9% homology for κ -CN promoter.

한우의 BoLA DRB3 exon2 유전자의 특성

강호범,류승희,이상훈,전병순,상병찬 충남대학교 농업과학연구소 1998 농업과학연구 Vol.25 No.1

This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

홀스타인종 유우의 혈청단백질 및 효소의 유전적 변이체

상병찬,류승희,서길웅,이창수 충남대학교 농업과학연구소 1995 농업과학연구 Vol.22 No.2

This study was carried out to examine the genetic constitution of serum proteins and enzymes in Holstein Friesian cattle population. The genetic variants of post-transferrin-2(pTf-2), transferrin(Tf), post-albumin(pAlb), ceruloplasmin(Cp) and amylase-I(Am-I) were analyzed by using PAGE(polyacrylamide gel electrophoresis) and STAGE(starch gel electrophoresis). In serum proteins, the pTf-2 locus were observed to be controlled by codominant alleles designated F and S, and the distribution of genotypes were 76.34, 14.50 and 9.10% for pTf-2 FF, FS and SS types, respectively. The gene frequencies of the pTf-2 F and S allele were 0.836 and 0.164. The Tf locus were found to be controlled by four alleles, Tf A, D1, D2 and E at a single locus, and the distribution of genotypes were 6.11, 32.06, 19.08, 1.53, 10.69, 18.32, 9.92 and 2.29% for Tf AA, AD1, AD2, AE, D1D1, D1D2, D2D2 and D2E type, respectively. The gene frequencies of the Tf A, D1, D2 and E wee 0.321, 0.359, 0.298 and 0.019. The pAlb locus were identified to be genetically controlled by two alleles, pAlb F and S allele, and the distribution of genotypes were 32.06, 29.77 and 38.17% for pAlb FF, FS and SS types, respectively. The gene frequencies of the pAlb F and S allele were 0.461 and 0.531. The Alb locus were observed to be controlled by Alb A and B allele, and the gene frequencies of these were 0.996 and 0.004. In serum enzymes, the Cp locus were found to be controlled by F and S allele, and the distribution of genotypes were 46.57, 27.48 and 25.95% for Cp FF, FS and SS types, respectively. The gene frequencies of F and S allele were 0.603 and 0.394. The Am-I locus were observed to be controlled by Am-I B and C allele, and the distribution of genotypes were 39.69, 21.73 and 38.93% for Am-I BB, BC and CC types, the gene frequencies of Am-I B and C were 0.503 and 0.497, respectively.

재래한우의 혈액단백질 및 효소의 유전적 다형현상

상병찬,류승희,이상훈,한성욱,이창수 충남대학교 농업과학연구소 1995 농업과학연구 Vol.22 No.2

This study was conducted to investigate the genetic constitution of blood proteins and enzymes in 238 Korean Native cattle reared at Korean Native Cattle Breeding Center, National Livestock Cooperative Federation. The genetic polymorphisms of post-transferrin-2(pTf-2), transferrin(Tf), post-albumin(pAlb), albumin(Alb), ceruloplamin(Cp), amylase-I(Am-I) and hemoglobin(Hb) were analyzed by using PAGE(polyacrylamide gel electrophoresis) and STAGE(starch gel electrophoresis). The genotypes and gene frequencies were estimated at these loci for each blood proteins and enzymes. The results obtained from this study were summarized as follows : 1. The pTf-2 locus were identified to be genetically controlled by codominant alleles designated pTf-2 F and S, and the distribution of genotypes were 46.22, 46.64 and 7.14% for pTf-2 FF, FS and SS types, and the gene frequencies of the pTf-2 F and S allele were 0,695 and 0.305, respectiveley. 2. The Tf locus were found to be controlled by Tf A, D1, D2 and E alleles, and the distributioin of genotypes were 0.84, 13.87, 13.03, 10.92, 22.27, 12.61, 2.94, 15.51, 6.72 and 1.68% for Tf AA, AD1, AD2, AE, D1D1, D1D2, D1E, D2E and EE types, and the gene frequencies of Tf A, D1, D2 and E were 0.197, 0.430, 0.191 and 0.081, respectively. 3. The pAlb locus were observed to be controlled by two alleles, pAlb F and S, and the distribution of genotypes were 42.86, 33.19 and 23.95% for pAlb FF, FS and SS types, and the gene frequencies were 0.595 and 0.405 for Tf F and S allele, respectively. Also the gene frequencies of Alb was 1.000 of Alb A allele. 4. The Cp locus were identified to be controlled by Cp F and S allele, and the distribution of genotypes were 23.11, 34.87 and 42.02% for Cp FF, FS and SS types, and the gene frequencies were 0.405 and 0.595 for Cp F and S allele, respectively. 5. The Am-I locus were observed to be genetically controlled by Am-I B and C allele, and the distribution of genotypes were 51.26, 16.81 and 31.92% for Am-I BB, BC and CC types, and the gene frequencies of Am-I B and C alleles were 0.597 and 0.403, respectively. 6. The Hb locus were found to be controlled by Hb A and B alleles, and the distribution of genotypes were 93.19, 16.39 and 0.42% for Hb AA, AB and BB types, and the gene frequencies of Hb A and B alleles were 0.914 and 0.086, respectively.

한우와 Holstein종의 κ -casein 유전자의 발현조절부위의 염기서열 분석

상병찬,류승희,이상훈,Sang, Byung Chan,Ryoo, Seung Heui,Lee, Sang Hoon 충남대학교 농업과학연구소 2000 농업과학연구 Vol.27 No.1

본 연구는 한우와 Holstein종의 $\kappa$ -CN 유전자 발현조절부위, 즉 $\kappa$ -CN gene 5'-flanking region에 대한 특성을 구명하기 위하여 specific primer를 이용하여 PCR(polymerase chain reaction) 방법으로 분석하였다. PCR 증폭 결과, 349bp 크기의 증폭산물을 얻었으며 한우와 Holstein종의 $\kappa$ -CN 유전자의 발현조절부위의 염기서열 분석결과에 있어서는 -82bp에서 -431bp까지 분석되었으며, 종간의 염기서열 비교에서 -386bp에서는 Holstein종에서는 T 염기가 한우에서는 C 염기로 치환되었으며, -241bp와 -192bp에서는 Holstein종에서는 T 및 C 염가가 존재하였으나, 한우에서는 결실되었다. 또한, -183bp에서는 한우에서는 T 염기가 존재하였으나, Holstein종에서는 염기가 결실되어 있음을 알 수 있었다. 또한 한우와 Holstein 종간의 상동성 분석에 있어서는 한우 $\kappa$ -CN 유전자 발현조절부위의 염기서열이 Holstein종과 98.9% 의 상동성을 보였다. This study was carried out to investigate the characterizations of $\kappa$ -CN genes of milk protein. In order to find out the characterizations of $\kappa$ -CN genes, the nucleotide sequences of 5'-flanking regions of $\kappa$ -CN genes were analyzed by PCR(polymerase chain reaction) technique with specific primers in order to investigate the characterizations of these promoters in Korean Native cattle and Holstein. PCR products obtained 349bp fragment and the nucleotide sequences of Korean Native cattle and Holstein of $\kappa$ -CN gene S'-flanking region was analyzed to -82bp from -431bp. On the comparison of each breed, Holstein substituted T$\rightarrow$C at -386bp, and -241bp(T) and -192bp(C) existed, but Korean Native cattle was deleted. Also, Korean Native cattle was existed T at -183bp but Holstein was not. The homology analysis of between Korean Native cattle and Holstein was showed 98.9% homology for $\kappa$ -CN promoter.

Evaluation of calving interval and selection indices in Korean native cows

Inchul Choi,Dooho Lee,Jong-Gwan Lee,Seung-Hwan Lee,Seung-Heui Ryoo 충남대학교 농업과학연구소 2020 Korean Journal of Agricultural Science Vol.47 No.3

Identification of SNPs in TG and EDG1 genes and their relationships with carcass traits in Korean cattle (Hanwoo)

Muhammad Cahyadi(카야디),Dyah Maharani(디아 마하라니),류승희(Seung Heui Ryoo),Seung Hwan Lee(이승환),Jun Heon Lee(이준헌) 충남대학교 농업과학연구소 2012 농업과학연구 Vol.39 No.3

Thyroglobulin (TG) gene was known to be regulated fat cell growth and differentiation and the endothelial differentiation sphingolipid G-protein-coupled receptor 1 (EDG1) gene involves blood vessel formation and known to be affecting carcass traits in beef cattle. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) in both TG and EDG1 genes and to analyze the association with carcass traits in Korean cattle (Hanwoo). The T354C SNP in TG gene located at the 3’ flanking region and c.-312A>G SNP located at 3’-UTR of EDG1 gene were used for genotyping the animals using PCR-RFLP method. Three genotypes were identified in T354C SNP in TG gene and only two AA and AG genotypes were observed for the c.-312A>G SNP in EDG1 gene. The results indicated that T354C SNP in TG gene was not significantly associated with carcass traits. However, the c.-312A>G SNP in EDG1 gene had significant effects on backfat thickness (BF) and yield index (YI). These results may provide valuable information for further candidate gene studies affecting carcass traits in Korean cattle and may use as marker assisted selection for improving the quality of meat in Hanwoo.