CAJ | 학술논문

为了研究NR5A2基因在乌骨绵羊中的序列特征,利用PCR和克隆测序技术获得乌骨绵羊NR5A2基因的外显子序列,用DNAMAN软件和DNAStar软件对序列进行比对和拼接、运用C1usta1软件与其他物种相应序列进行同源比对分析,MEGA5.1软件用来构建哺乳动物NR5A2蛋白质系统进化树,用DNA池方法对其编码区序列的SNPs位点进行筛选.结果显示：乌骨绵羊 NR5A2基因编码区全长1488 bp,共编码495个氨基酸；乌骨绵羊的NR5A2基因编码区核苷酸序列与湖羊和牛的同源性分别为99.87%和85.51%；氨基酸序列的一致性分别为99.41%和89.66%.与湖羊相比,乌骨绵羊 NR5A2基因编码区共发现3个单碱基突变( C1069A、C1419T 和G1485A),其中C1069A位点突变引起了氨基酸的改变,从亮氨酸变为异亮氨酸.在乌骨绵羊的编码区现了一个单核苷酸突变位点G999C.
위료연구NR5A2기인재오골면양중적서렬특정,이용PCR화극륭측서기술획득오골면양NR5A2기인적외현자서렬,용DNAMAN연건화DNAStar연건대서렬진행비대화병접、운용C1usta1연건여기타물충상응서렬진행동원비대분석,MEGA5.1연건용래구건포유동물NR5A2단백질계통진화수,용DNA지방법대기편마구서렬적SNPs위점진행사선.결과현시：오골면양 NR5A2기인편마구전장1488 bp,공편마495개안기산；오골면양적NR5A2기인편마구핵감산서렬여호양화우적동원성분별위99.87%화85.51%；안기산서렬적일치성분별위99.41%화89.66%.여호양상비,오골면양 NR5A2기인편마구공발현3개단감기돌변( C1069A、C1419T 화G1485A),기중C1069A위점돌변인기료안기산적개변,종량안산변위이량안산.재오골면양적편마구현료일개단핵감산돌변위점G999C.
To ana1yze the sequence characterization of NR5A2 gene in b1ackbone sheep, exon sequences of NR5A2 were obtained using PCR and c1oning and sequencing techno1ogy, assemb1ed in turn using DNAStar and DNAMAN software and then b1asted with its corresponding sequences of other species with C1usta1 software. The fu11 1ength of coding region of NR5A2 was 1 488 bp in b1ackbone sheep encoding 495 amino acids. The NR5A2 coding region shared 99. 87% and 85. 51% simi1arities in nuc1eotide sequence with Hu sheep and catt1e, respective1y, and 99. 41% and 89. 66% simi1arities in amino acids sequence, respective1y. The phy1ogenetic tree constructed based on NR5A2 protein using MEGA5. 1 software revea1ed b1ackbone sheep and Hu sheep were c1ustered c1ose1y together, and then formed a c1uster with catt1e. B1ackbone sheep, Hu sheep and catt1e exhibited a c1oser genetic re1ation to 1ivestock, such as swine, horse, and dog than primate. B1ackbone sheep was distinct from pou1ty and fishery. Compared with Hu sheep, three sing1e nuc1eotide po1ymorphism(C1069A, C1419T and G1485A) in coding region of NR5A2 of b1ackbone sheep were detected, among which, C1069A mutation caused the changes of amino acid, from 1eucine to iso1eucine. A SNP site, G999C, was found in the coding region of b1ackbone sheep using DNA poo1.