摘要
本研究旨在克隆小尾寒羊B细胞易位基因2(B-cell Translocation Gene 2,BTG2)基因编码CDs区,并对其进行生物信息学分析,同时检测该基因在羔羊期各组织中的mRNA表达情况。通过RT-PCR和TA克隆方法获得BTG2基因CDs区序列;利用在线软件对该基因进行生物信息分析;采用qPCR检测该基因在3、40日龄绵羊的各组织中的表达情况。结果表明:成功获得小尾寒羊BTG2基因CDs区序列453 bp;绵羊与牛、猪、虎鲸、人、大鼠、小鼠同源性分别为98.9%、90.29%、94.04%、88.52%和85.65%、86.98%,且绵羊与牛的亲缘关系最近,与小鼠的亲缘关系最远。理化性质结果显示BTG2基因编码150个氨基酸,蛋白分子式为C748H1190N208O213S8,分子质量为17 ku,理论等电点(pI)为9.14,半衰期为30 h,属于不稳定亲水性蛋白,且不存在跨膜结构和信号肽序列,蛋白结构主要通过α-螺旋、无规则卷曲和β股组成,为混合型蛋白;qPCR检测到BTG2基因mRNA在不同日龄羔羊心脏、肝脏、脾脏、肺脏、肾脏、肠和肌肉均有表达,且均在肝脏中表达量最高;在心脏、肝脏、脾脏、肺脏组织中,40日龄BTG2表达量极显著高于3日龄,而在肾脏、肠和肌肉中,40日龄极显著低于3日龄。结果表明,BTG2广泛存在各组织中,且随着日龄增加,在各组织中的表达趋势不同,提示其功能的重要性和多重性,并且BTG2在肝组织中表达量最高,说明BTG2可能与脂代谢以及糖代谢等调控功能相关。
The purpose of this study was to clone and analyze CDs region of B-cell translocation gene 2(BTG2)of Smalltail Han sheep by bioinformatics,and detect the mRNA expression profile of the gene in different tissues at lambs stages.The CDs region sequence of BTG2 gene was obtained by RT-PCR and TA cloning.The online software was used to analyze the biological information of BTG2 gene.qPCR was used to detect the expression trend of this gene in different tissues of 3-day and 40-day sheep.The results showed that the CDs sequence of BTG2 gene in Small-tail Han sheep was obtained successfully in this paper.The homology of ovisaries,Bostaurus,Susscrofa,Orcinusorca,Homo sapiens,Rattusnorvegicus and Musmusculus were 98.9%,90.29%,94.04%,88.52%,85.65%and 86.98%,respectively.Sheep are the closest relatives of cattle and the furthest relatives of mice.The physical and chemical properties showed BTG2 gene encoding 150 amino acids and protein formula was C748H1190N208O213S8 with a molecular weight of 17 ku,theoretical isoelectric point(pI)of 9.14 and a half-life of 30h,belongs to the unstable hydrophilic protein.There is no transmembrane structure and signal peptide sequence in BTG2,and the protein structure is mainly composed of helical,irregular crimp and lamina,and it is a hybrid protein.The BTG2 gene was found to be expressed in the heart,liver,spleen,lungs,kidneys,intestines and muscles of lambs at different age by qPCR.The expression of BTG2 gene is the highest at different lambs age,respectively.Further analysis revealed that the BTG2 expression level of 40-day-old sheep was significantly higher than that of 3-day-old sheep in the tissues of heart,liver sheep,spleen and lung(P<0.001),while the BTG2 expression level of 40-day-old sheep in the tissues of kidney,intestine and muscle was significantly lower than that of 3-day-old sheep(P<0.001).The results showed that BTG2 was widely used in various organizations.With the increase of age,the expression trend of BTG2 in each tissue was different,indicating the importance and mul
作者
高一
刘理想
肖成
金海国
马惠海
薛佳佳
魏天
曹阳
GAO Yi;LIU Lixiang;XIAO Cheng;JIN Haiguo;MA Huihai;XUE Jiajia;WEI Tian;CAO Yang(Institute of Animal Biotechnology,Jilin Academy of Agricultural Science,Jilin Gongzhuling 136100,China;College of Animal Science and Technology,Jilin Agricultural University,Jilin Changchun 130118,China)
出处
《中国畜牧杂志》
CAS
北大核心
2021年第2期74-79,共6页
Chinese Journal of Animal Science
基金
吉林省农业科学院创新工程资助(CXGC2017ZD002)
国家肉羊产业技术体系(CARS38)。
