山羊FTL基因克隆、序列分析和组织表达研究  

Study on cloning,sequence analysis and tissue expression of FTL gene in goats

作　　者：龚静 曾献春[2] GONG Jing;ZENG Xianchun(Callrge of Agric ulure and Frestry Health,Sichuan Open University,Chengdu 610073,China;School of Labuatory Medic ine,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]四川开放大学农林卫生学院,成都610073 [2]成都医学院检验医学院,成都610500

出　　处：《黑龙江畜牧兽医》2022年第15期58-63,136,137,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：山羊溶菌酶的抗逆性改造项目“2021年青藏高原动物遗传资源保护与利用教育部实验室对外发布项目”。

摘　　要：为了探讨山羊体内铁蛋白轻链(ferritin light chain,FTL)的结构及生物学功能,试验利用RT-PCR技术克隆得到山羊FTL基因序列,通过在线软件分析其生物学特性,构建物种进化树揭示物种间亲缘关系;根据克隆获得的FTL基因序列,通过实时荧光定量PCR技术检测山羊FTL基因在不同组织中表达水平;复苏前期保存的山羊肌内前体脂肪细胞,利用实时荧光定量PCR技术检测FTL基因在诱导分化各时间点的相对表达量。结果表明:克隆获得的山羊FTL基因编码区(CDS)序列全长为745 bp,其中包括完整的大小为528 bp的ORF,共编码173个氨基酸;FTL蛋白有6个丝氨酸磷酸化位点,4个苏氨酸磷酸化位点,有23个带正电荷的赖氨酸和精氨酸残基,26个带负电荷的谷氨酸和天冬氨酸残基,无跨膜结构域;山羊FTL基因与牛进化亲缘关系最近;FTL基因在山羊体内各器官/组织中均有不同程度的表达,其中在肝脏中的相对表达量最高;FTL基因在24小时时相对表达量最高,24 h后的表达量呈不断下降的趋势。说明山羊FTL蛋白与脂肪代谢具有一定直接或间接关系。In order to explore the structure and biological function of ferritin light chain(FTL)in goats,the FTL gene sequence in goats was cloned by RT-PCR.Its biological characteristics were analyzed by online software,and the species evolutionary tree was constructed to reveal the genetic relationship between species.According to the cloned FTL gene sequence,the expression level of goat FTL gene in different tissues was detected by real-time fluorescence quantitative PCR(qRT-PCR).Previously preserved intramuscular preadipocytes of goats were resuscitated,and the expression level of FTL gene at each time point of induction and differentiation was assessed by qRT-PCR.The results showed that the CDS of goat FTL gene was 745 bp,including complete ORF 528 bp,encoding 173 amino acids.FTL protein had 6 serine phosphorylation sites(SER)and 4 threonine phosphorylation sites(THR),23 positively charged lysine and arginine residues,26 negatively charged glutamate and aspartic acid residues,and no transmembrane domain.The genetic relationship between goat FTL gene and cattle was the closest.FTL gene was expressed in various tissues of goats,and the relative expression was the highest in liver.The relative expression of FTL gene was the highest at 24 h,and the expression showed a declining trend after 24 h.These results indicated that goat FTL protein was directly or indirectly related to fat metabolisms.

关 键 词：山羊 FTL基因 克隆 组织表达 时序表达 

分 类 号：S827[农业科学—畜牧学] Q785[农业科学—畜牧兽医]