CAJ | 학술논문

为了获得湖羊UCP3基因编码区全序列并分析其序列特征,同时研究其在湖羊不同部位肌肉中的表达变化,选取初生与1、2、3、4、5和6月龄湖羊公羔各5只,提取屠宰后背最长肌、腰大肌、前腿肱二头肌和后腿股二头肌RNA,用TA克隆技术获得湖羊UCP3基因编码区序列,用DNAMAN软件分析UCP3基因在哺乳动物间的保守性,同时基于UCP3蛋白质序列用MEGA5.1软件构建系统发育树,用Real-time PCR检测湖羊不同肌肉部位UCP3基因的表达水平并分析其在不同发育时期的变化。结果显示：湖羊UCP3基因编码区全长936 bp,编码311个氨基酸残基,在哺乳动物中相对保守。根据UCP3蛋白质氨基酸序列构建的系统发育树显示湖羊与牛的亲缘关系最近。Real-time PCR结果显示,初生至6月龄的湖羊不同部位肌肉UCP3基因的表达模式基本相似,即在出生时有较低的水平,然后上升,随着月龄的增加又下降,但UCP3基因在后腿股二头肌中的表达量整体偏低,在腰大肌中表达水平变化较大,3月龄的表达水平显著高于初生和5月龄。
위료획득호양UCP3기인편마구전서렬병분석기서렬특정,동시연구기재호양불동부위기육중적표체변화,선취초생여1、2、3、4、5화6월령호양공고각5지,제취도재후배최장기、요대기、전퇴굉이두기화후퇴고이두기RNA,용TA극륭기술획득호양UCP3기인편마구서렬,용DNAMAN연건분석UCP3기인재포유동물간적보수성,동시기우UCP3단백질서렬용MEGA5.1연건구건계통발육수,용Real-time PCR검측호양불동기육부위UCP3기인적표체수평병분석기재불동발육시기적변화。결과현시：호양UCP3기인편마구전장936 bp,편마311개안기산잔기,재포유동물중상대보수。근거UCP3단백질안기산서렬구건적계통발육수현시호양여우적친연관계최근。Real-time PCR결과현시,초생지6월령적호양불동부위기육UCP3기인적표체모식기본상사,즉재출생시유교저적수평,연후상승,수착월령적증가우하강,단UCP3기인재후퇴고이두기중적표체량정체편저,재요대기중표체수평변화교대,3월령적표체수평현저고우초생화5월령。
This study aims to obtain the full coding region of UCP3 gene in Hu sheep, characterize its sequence and investigate the mRNA level in sheep muscles. Each five male Hu lamb at 0 to 6 months ages were chosen and slaughtered, and total RNA from longissimus muscles, psoas muscles and front and rear leg biceps femoris muscles were extracted. The coding region of UCP3 gene of Hu sheep was cloned using TA cloning technoloty, and its conservation among different mam-mals was analyzed using DNAMAN software. Its mRNA level of different muscles of Hu sheep was investigated using real time PCR. The results showed that the coding region of UCP3 gene in Hu sheep was 936 bp, encoding 311 amino acid residues, and was relatively conserved in mammals. Phylogenetic tree constructed based on protein sequence of UCP3 showed that the genetic relationship between Hu sheep and cattle was the closest. Real-time PCR results revealed that UCP3 in different parts of muscles of differently-aged Hu sheep shared similar mRNA expression patterns, which were low at birth, and went up slightly and down afterwards over age. The expression level of UCP3 gene was relatively low in the rear leg biceps femoris muscles, and fluctuated in psoas muscles. UCP3 gene showed higher levels in 3-mouth-old sheep than those in the newborns and 5-mouth-old sheep.