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돼지 유전체 염기서열을 이용한 내인성 리트로 바이러스 분석에 관한 연구
유성란(Seong-Lan Yu),이준헌(Jun Heon Lee) 충남대학교 농업과학연구소 2009 Korean Journal of Agricultural Science Vol.36 No.2
본 연구는 현재까지 발표된 돼지의 genomic sequence 정보를 이용하여 PERV들의 정확한 삽입 위치를 파악하고 그들의 특성을 분석하고자 실시하였으며 총 2.7 Gb인 돼지 genome 염기서열 중 4.2%인 114 Mb의 염기서열에서 PERV sequence를 확인한 결과 총 8개의 PERV sequence를 확인할 수 있었다. 확인된 PERV sequence중 7개는 유전자내에 deletion이 확인되었으며 나머지 한 개의 PERV도 gag와 env 유전자에 stop codon이 확인되어 정상적인 PERV로 발현되지 않을 것으로 추정되었다. 본 연구는 돼지를 이용한 이종장기이식과 관련하여 PERV를 제어하기 위한 중요한 기초 연구 자료를 제공할 것으로 사료된다. This study was conducted to identify the PERV (Porcine Endogenous Retrovirus) integration sites and their characterizations using the porcine genomic sequence information. Total 114 Mb (4.2%) sequence of the 2.7 Gb pig genome was investigated for the PERV sequences. As the results, 8 PERV sequences were identified and their genomic structures were deduced from the BLAST searches against previously known PERV genes. Seven PERVs have internal deletions in the protein coding region and they will not be functional. The other one also has internal deletions in the gag and env genes, indicating this PERV is also defective. Even though we could not identify the functional PERVs in this study, the results presented here can be used for the fundamental research materials for controlling PERV infections in relation to xenotransplantation using porcine organs and tissues.
돼지 유전체 염기서열을 이용한 내인성 리트로 바이러스 분석에 관한 연구
유성란(Seong-Lan Yu),이준헌(Jun Heon Lee) 충남대학교 농업과학연구소 2009 농업과학연구 Vol.36 No.2
본 연구는 현재까지 발표된 돼지의 genomic sequence 정보를 이용하여 PERV들의 정확한 삽입 위치를 파악하고 그들의 특성을 분석하고자 실시하였으며 총 2.7 Gb인 돼지 genome 염기서열 중 4.2%인 114 Mb의 염기서열에서 PERV sequence를 확인한 결과 총 8개의 PERV sequence를 확인할 수 있었다. 확인된 PERV sequence중 7개는 유전자내에 deletion이 확인되었으며 나머지 한 개의 PERV도 gag와 env 유전자에 stop codon이 확인되어 정상적인 PERV로 발현되지 않을 것으로 추정되었다. 본 연구는 돼지를 이용한 이종장기이식과 관련하여 PERV를 제어하기 위한 중요한 기초 연구 자료를 제공할 것으로 사료된다. This study was conducted to identify the PERV (Porcine Endogenous Retrovirus) integration sites and their characterizations using the porcine genomic sequence information. Total 114 Mb (4.2%) sequence of the 2.7 Gb pig genome was investigated for the PERV sequences. As the results, 8 PERV sequences were identified and their genomic structures were deduced from the BLAST searches against previously known PERV genes. Seven PERVs have internal deletions in the protein coding region and they will not be functional. The other one also has internal deletions in the gag and env genes, indicating this PERV is also defective. Even though we could not identify the functional PERVs in this study, the results presented here can be used for the fundamental research materials for controlling PERV infections in relation to xenotransplantation using porcine organs and tissues.
한국형 홀스타인종 젖소의 BLV env 유전자의 특성분석
정행진(Hang-Jin Jeong),유성란(Seong-Lan Yu),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),서국현(Guk-Hyun Suh),류승희(Seung-Heui Ryoo),정상일(Sang-Il Chung),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2011 농업과학연구 Vol.38 No.2
This study was performed to investigate the characterization of infectious BLV env gene isolated form Korean Holstein Cattle and to determine its incoming origin. Gp51 region of BLV env gene known as having important role in immunological function was characterized using PCR-RFLP sequencing and phylogenetic analysis. BLV env gene was grouped into PCR-RFLP patterns with three restriction endonucleases including Pvu Ⅱ, BamHI and HaeⅢ, and we identified two new RFLP patterns from nucleotide sequences of each group. Phylogenetic analysis showed that 80% of the Korean Holstein was included in the USA and Japanese group. These results here can provide a valuable information about the character of the BLV env gene and research on infection route of BLV.
한국형 홀스타인종 젖소의 BLV env 유전자의 특성분석
정행진(Hang-Jin Jeong),유성란(Seong-Lan Yu),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),서국현(Guk-Hyun Suh),류승희(Seung-Heui Ryoo),정상일(Sang-Il Chung),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2011 Korean Journal of Agricultural Science Vol.38 No.2
This study was performed to investigate the characterization of infectious BLV env gene isolated form Korean Holstein Cattle and to determine its incoming origin. Gp51 region of BLV env gene known as having important role in immunological function was characterized using PCR-RFLP sequencing and phylogenetic analysis. BLV env gene was grouped into PCR-RFLP patterns with three restriction endonucleases including Pvu Ⅱ, BamHI and HaeⅢ, and we identified two new RFLP patterns from nucleotide sequences of each group. Phylogenetic analysis showed that 80% of the Korean Holstein was included in the USA and Japanese group. These results here can provide a valuable information about the character of the BLV env gene and research on infection route of BLV.
홀스타인종 젖소에 있어서 PCR과 ELISA기법을 이용한 BLV 감염진단
정행진(Hang-Jin Jeong),유성란(Seong-Lan Yu),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),서국현(Guk-Hyun Shu),류승희(Seung-Heui Ryoo),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2011 농업과학연구 Vol.38 No.1
This study was conducted to investigate the farm situation about bovine leukemia virus(BLV) infection that greatly influence productivity in dairy cattle and compare the accuracy of diagnosis for BLV infection between PCR and ELISA techniques. Blood samples of 193 heads from 5 herds in Chungnam and Chungbuk area were used to analyze BLV gene and serum, and the results were obtained as follows. The amplified BLV gene in dairy cattle by PCR technique resulted in 226 bp, 596 bp and 434 bp, respectively, for gag, pol and env, which were well amplified. The infection rates of BLV virus diagnosed by PCR and ELISA techniques ranged from 80.55 to 100% and from 22.22 to 86.95%, respectively, and the infection rates among 5 herds were significantly different in both methods (P<0.05). Further, the average infection rates of 5 herds were 87.05 and 63.21%, respectively, for PCR and ELISA techniques. Kappa statistics for examining consistency of diagnosis by PCR and ELISA techniques showed 0.246, which represents low consistency. Consequently, PCR based BLV technique was considered as a corrective measure for diagnosis of BLV infection in Holstein dairy cattle.
홀스타인종과 한우에 있어서 BoLA-DRB3 유전자의 단일염기다형과 반수체 분석
정행진(Hang-Jin Jeong),유성란(Seong-Lan Yu),라세둘(M. R. Hoque),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),류승희(Seung-Heui Ryoo),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2011 농업과학연구 Vol.38 No.1
BoLA (bovine leukocyte antigens) have been known as gene complex related with bovine diseases and immunological traits. This study was conducted to find out the characteristics of BoLA-DRB3 gene related to mastitis and BL(bovine leukocyte) from 280 cattle [193 animals of Holstein cattle and 87 animals of Hanwoo]. As a result, five PCR-RFLP types (b, d, e, f and g) using HaeⅢ restriction enzyme, three BstYI restriction patterns (b, d and e) and eight RsaI restriction types(b, d, f, I, j, n, o and w) were identified. Moreover, we identified new d' type (197→175/22), having one more cutting site by BstYI enzyme than d type allele and n' type (180→169/11) having one more cutting site by RsaI enzyme than n allele was additionally identified. Next, we identified 53 SNPs in BoLA-DRB3 exon2 from 280 cattle. SNP frequency and heterozygosity of Holstein and Hanwoo were investigated in all the SNP genotype. These results might be based on research for identifying marker associated with bovine diseases.
한국 Holstein종 유우집단의 DGAT1 유전자의 특성분석
손지영(Ji-Young Son),정행진(Hang Jin-Jeong),유성란(Seong-Lan Yu),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),류승희(Seung-Heui Ryoo),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2009 농업과학연구 Vol.36 No.2
본 연구는 한국형 Holstein종 젖소집단의 DGAT1 유전자의 특성을 구명하고, DGAT1의 유전적 다형과 산유형질인 유량 및 유지량 간의 연관성을 구명하여 젖소집단의 유전적 개량을 위한 분자유전학적 접목을 위하여 실험을 실시하였다. Holstein종의 genomic DNA로부터 PCR기법을 이용하여 DGAT1 유전자좌를 specific primers로 증폭한 후, 1.5% agarose gel에 전기영동한 결과 411 bp의 단편이 양호하게 증폭되었음을 확인하였다. DGAT1 유전자의 염기서열을 분석한 결과 DGAT1 Q 대립유전자의 216-218 bp의 염기서열이 AUG(lysine, K)였으나, 동위치의 대립유전자 q의 염기서열은 GCG(alanine, A)로 치환되어 있음을 확인할 수 있었다. 한편 한국 Holstein종과 NCBI에서 보고된 bovine DGAT1 유전자 단편의 염기서열간에는 100% 상동성을 보였다. 한국 Holstein종 유우집단의 DGAT1 유전자형의 분포는 DGAT1 QQ, Qq 및 qq 유전자 분포가 각각 16.43, 36.43 및 47.14%로 qq 유전자형빈도가 다른 유전자형에 비하여 높았으며, 유전자빈도는 DGAT1 Q 및 q 빈도가 각각 0.35 및 0.65로 q의 빈도가 높았다. DGAT1 유전자형과 산유량인 유량 및 유지량간의 연관성에 있어서는 DGAT1 유전자형이 유량 및 유지량에서 유의적인 차이(P<0.05)를 보였으며, DGAT1 Qq 유전자형이 QQ 및 qq 유전자형에 비하여 유량과 유지량에서 유의적인 차이(P<0.05)로 높은 수치를 나타냈다. This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).
한국 Holstein종 유우집단의 DGAT1 유전자의 특성분석
손지영(Ji-Young Son),정행진(Hang Jin-Jeong),유성란(Seong-Lan Yu),이준헌(Jun-Heon Lee),도창희(Chang-Hee Do),류승희(Seung-Heui Ryoo),상병찬(Byung-Chan Sang) 충남대학교 농업과학연구소 2009 Korean Journal of Agricultural Science Vol.36 No.2
본 연구는 한국형 Holstein종 젖소집단의 DGAT1 유전자의 특성을 구명하고, DGAT1의 유전적 다형과 산유형질인 유량 및 유지량 간의 연관성을 구명하여 젖소집단의 유전적 개량을 위한 분자유전학적 접목을 위하여 실험을 실시하였다. Holstein종의 genomic DNA로부터 PCR기법을 이용하여 DGAT1 유전자좌를 specific primers로 증폭한 후, 1.5% agarose gel에 전기영동한 결과 411 bp의 단편이 양호하게 증폭되었음을 확인하였다. DGAT1 유전자의 염기서열을 분석한 결과 DGAT1 Q 대립유전자의 216-218 bp의 염기서열이 AUG(lysine, K)였으나, 동위치의 대립유전자 q의 염기서열은 GCG(alanine, A)로 치환되어 있음을 확인할 수 있었다. 한편 한국 Holstein종과 NCBI에서 보고된 bovine DGAT1 유전자 단편의 염기서열간에는 100% 상동성을 보였다. 한국 Holstein종 유우집단의 DGAT1 유전자형의 분포는 DGAT1 QQ, Qq 및 qq 유전자 분포가 각각 16.43, 36.43 및 47.14%로 qq 유전자형빈도가 다른 유전자형에 비하여 높았으며, 유전자빈도는 DGAT1 Q 및 q 빈도가 각각 0.35 및 0.65로 q의 빈도가 높았다. DGAT1 유전자형과 산유량인 유량 및 유지량간의 연관성에 있어서는 DGAT1 유전자형이 유량 및 유지량에서 유의적인 차이(P<0.05)를 보였으며, DGAT1 Qq 유전자형이 QQ 및 qq 유전자형에 비하여 유량과 유지량에서 유의적인 차이(P<0.05)로 높은 수치를 나타냈다. This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).