山羊Δfosb基因cDNA的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Dairy Goat Δfosb cDNA

克隆山羊Δfosb基因的cDNA片段,分析其基因序列,为进一步研究Δfosb基因在奶山羊钙代谢机制中的作用提供理论依据。无菌采集泌乳期奶山羊乳腺组织样并提取总RNA备用。根据已知其他物种的fosb基因保守性区域设计特异性引物,采用RT-PCR方法扩增奶山羊Δfosb基因的CDS区,测序后进行核苷酸和推导的氨基酸序列分析,并利用保守区序列对9个物种进行聚类分析。结果显示,得到953 bp的Δfosb基因cDNA片段(GenBank登录号:FJ455501)。核酸序列分析结果显示,该基因编码240个氨基酸,与人、小鼠和牛的核苷酸序列比对,同源性分别为93.33%、88.19%和98.47%,氨基酸同源性分别为96.68%、95.83%和99.17%。表明该基因在不同物种间具有高度的保守性。9个物种Δfosb基因的保守区聚类结果显示,羊和牛亲缘关系最近,其次依次为大鼠和小鼠、人和黑猩猩、马、狗、猪。

To clone the cDNA sequences of△fosb gene of dairy goat and to know its biological nature. Collecting the mammary gland tissue of goat during lactation under sterile conditions and extracting the total RNA was carried out. According to the conserved regions of fosb gene of other known species, a pair of specific primers were designed and the RT-PCR was performed to amplify the complete CDS sequences of △fosb from dairy goat. A fragment of 953bp was obtained through RT-PCR, which is with the GenBank accession No. FJ455501. The gene encoded for 240 amino acid residues. The similarity of goat △fosb cDNA sequence with that of human, mouse and cattle were determined as 93.33%, 88.19% and 98.47%, respectively,and the similarity of its amino acid sequence with that of human, mouse and cattle were determined as 96.68%, 95.83% and 99.17%, respectively. Phyloge- netic tree constructed from the conserved △fosb cDNA sequences of 9 different species illustrated a unique branches for goat and cattle, mouse and rat, human and chimpanzee, horse, dog, and pig. Goat and cattle grouped together first.