한우 myostatin 유전자의 SNP 및 발현분석

유성란,정기철,상병찬,이준헌 충남대학교 농업과학연구소 2004 농업과학연구 Vol.31 No.2

Myostatin is a transforming growth and differentiation factor-β family member that acts as a negative regulator of muscle growth. Previously, mutations in the myostatin gene were known to be related to double muscling phenotypes in cattle. Because myostatin gene is highly related to muscle mass, also meat quality, in cattle, we sequenced whole myostatin mRNA and investigated the SNPs (Single Nucleotide Polymorphisms) in Korean cattle (Hanwoo). The results indicated that Hanwoo had an SNP in nt2385 and this mutation can be a useful marker with further verifications. We also investigated expression patterns of the myostatin gene from various muscle tissues and organs. Northern blotting results indicated that myostatin expression was restricted in muscles with variable expression levels. The results presented here can be used as a valuable information for meat quality related traits and muscle mass in cattle.

한우 c-fos 유전자의 염기서열 및 발현분석

유성란,정행진,정기철,이준헌,조규완,최재관,나기준,상병찬 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.6

Cellular FOS(c-fos) protein is a transcription factor that forms heterodimers mostly with c-jun family and stimulates the transcription of genes containing AP-1 regulatory elements. This c-fos expression can control growth and differentiation of various precursor cells including myoblasts. The controls by c-fos gene have been identified for affecting skeletal muscle fiber traits which are the key determinants of meat quality in pigs. As a first step for identifying the relationship between c-fos gene and meat quality traits in cattle, we fully sequenced 1,443 bp of Hanwoo c-fos mRNA and analyzed expression patterns from various organs and muscle tissues. The sequence identities of Hanwoo c-fos with that of human, pig and mouse showed 89.8%, 93.3% and rib muscle from 7 organs and 9 different parts of muscles investigated. These results presented here can be used as a valuable marker for meat quality related traits in cattle with further verification.

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.

유전 및 육종 소의 경제형질 관련 후보 유전자 및 Microarray 연구현황

유성란 ( S. L. Yu ),이준헌 ( J. H. Lee ) 한국축산학회 2006 한국축산학회지 Vol.48 No.2

소의 체세포핵이식태반과 정상태반간의 차등 발현 유전자 분석

유성란 ( Seong Lan Yu ),정행진 ( Hang Jin Jeong ),상병찬 ( Byung Chan Sang ),류승희 ( Seung Heui Ryoo ),정기철 ( Kie Chul Jung ),윤종택 ( Jong Taek Yoon ),성환후 ( Hwan Hoo Seong ),진동일 ( Dong Il Jin ),이준헌 ( Jun Heon Lee ) 한국동물자원과학회 2008 한국축산학회지 Vol.50 No.5

황갈색 재래닭의 간에서 성장 단계별 차등 발현 유전자 분석

이경연,유성란,정기철,장병귀,최강덕,이준헌,Lee, K.Y.,Yu, S.L.,Jung, K.C.,Jang, B.K.,Choi, K.D.,Lee, J.H. 한국가금학회 2007 韓國家禽學會誌 Vol.34 No.2

한국 재래닭에서 성장에 따른 유전자들의 발현 변화를 알아보고 성장 촉진, 대사 및 면역 관련 유전자를 발굴하기 위하여 주령별로 닭의 간에서 RNA를 추출하였으며 10개의 arbitrary ACPs를 이용하여 차등 발현되는 유전자를 조사하였다. 발현량에 현저한 차이를 보이는 5개의 유전자들이 선별되었으며, 이 중 3개의 유전자들은 BLAST search 결과 이미 기능이 알려진 FTH1, SAA와 HSP90B1으로 밝혀졌다. 그러나 2개의 유전자들은 닭의 genome sequence가 끝났음에도 불구하고 기능이 밝혀져 있지 않아 앞으로 이 유전자들의 기능에 대한 연구가 지속되어야 함을 의미한다. 본 연구에서 닭의 간에서 성장 단계별로 발현 차이를 보이는 유전자들은 앞으로 다른 유전자와 단백질들과의 관계를 통하여 닭의 성장 및 지방 대사를 이해하는데 도움을 줄 것으로 사료된다. The chicken liver has been involved in various biological functions including detoxification, glycogen storage and plasma protein synthesis. The aim of this study was to investigate differentially expressed genes in chicken liver in four different growing stages. Using 10 arbitrary Annealing Control Primers (ACPs), five differentially expressed genes have been identified. Based on the Basic Local Alignment Search Tool (BLAST) search results, three of them were matched with previously known genes, and the other two were matched with unknown EST sequence and a hypothetical protein, respectively. In order to confirm the expression results, quantitative real-time PCR was also performed. The high similarities between the expression data using arbitrary ACPs and quantitative real-time PCR indicate that the identified genes are the real differentially expressed genes in different growing stages. The genes identified in this study can be used as valuable biomarkers in chicken with further investigation of the functions.

한우 Band 3 및 CHS 유전병의 분자유전학적 기초연구

정행진,유성란,상병찬,이준헌 충남대학교 농업과학연구소 2005 농업과학연구 Vol.32 No.1

Many bovine genetic diseases are currently unidentified in Korea because of the relatively low monitoring systems in the livestock farms. The molecular detection system using PCR-RFLP of two genetic diseases, namely Band 3 (Erythrocyte Membrane Protein Band Ⅲ) and CHS (Chediak-Higashi Syndrome), have been identified in Japan and used for screening large number of cattle whether each individual has the genetic disease or not. Using the 22 unrelated Korean cattle (Hanwoo) individuals, molecular detection system based on PCR-RFLP have been investigated, which can be distinguishable carriers for the genetic diseases. Even though we could not found the causative mutations for two genetic diseases, the PCR-RFLP techniques used in this study are very valuable for the screening the genetics diseases in Korean cattle, especially for the proven or candidate bulls.

Expression analysis of Porcine Endogenous Retroviruses (PERVs) in Korean native pig organs

Oh, Hyung Gil,Jung, Woo Young,Yu, Seung Lan,이준헌 충남대학교 농업과학연구소 2011 농업과학연구 Vol.38 No.1

Pigs have anatomically and physiologically very similar to human and because of this, pigs are the possible xenotransplantation donors for human organs. PERVs (Porcine Endogenous Retroviruses) are known to be one of the possible obstacles for using porcine organs regardless of the immunological barriers. In order to understand the expression patterns of PERVs in Korean native pigs, we investigated PERV expressions in porcine liver, heart, spleen, and lung samples. After RNA extraction, two types of specific PERV envelope genes (ENV-A and ENV-B) were amplified using specific primers by RT-PCR. The results indicated that the variable PERV expressions were observed in inconsistent patterns among animals and tissues. The PERV expressions were verified with semi-quantitative real-time PCR with three replicates. Even though, these results confirm the previous findings that the PERVs were differentially expressed between animals and tissues. These results also give some valuable information for xenotransplantation when using the Korean native pigs as the organ donor.

재래산양 락토페린 유전자 5' Flanking 영역의 특성에 관한 연구

상병찬,유대열,유성란,이태훈,신상태,이경광 한국축산학회 2000 한국축산학회지 Vol.42 No.1

Lactoferrin(Lf), a ferric ion(Fe^(3+))-binding glycoprotein, is found most notably in milk and some exocrine secretions. It is a main antimicrobial component of colostrum and milk that contributes to protection of the infant against infectious diseases. To analyze 5' flanking region of Korean native goat lactoferrin gene, we cloned and sequenced 975 by of 5' flanking region of Korean native goat Lf gene. When compared to those of bovine and human Lf, the nucleotide sequences exhibited about 90.9% and 49.7% homology, respectively. The transcription start site was localized by primer extension to residue C, which is 38 nucleotides upstream from the ATG start codon. The 5' flanking region of Korean native goat lactoferrin gene contains putative transcription factor binding sites such as SP1, CAT, C/EBP and GATA-1.

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.