藏绵羊KAP3.2基因的cDNA克隆、序列分析及组织表达的研究

Cloning and tissue expression of KAP3.2 gene in Tibetan sheep

为研究藏绵羊角蛋白相关蛋白(KAP3.2)基因结构与功能,揭示该基因的组织特异性表达规律.以藏绵羊为研究对象,分别提取藏绵羊心、肝、脾、肾及皮肤组织中总RNA,并以此为模板,通过RT-PCR技术对藏绵羊KAP3.2基因的c DNA进行克隆、序列分析;利用Real-time PCR技术进行组织表达研究.结果表明:藏绵羊KAP3.2基因编码区全长297bp,编码98个氨基酸;藏绵羊与普通绵羊、山羊、藏羚羊、草原鼠、家鼠、人的相应核苷酸同源性分别为99%、97%、96%、79%、75%、73%;藏绵羊KAP3.2基因在皮肤中高表达,而在其它组织中相对表达量较低,说明该基因的表达具有组织特异性.

This study was carried out to obtain and analyze sequence of Tibetan sheep KAP3.2 gene by RT-PCR, and to determine its m RNA expression in different tissues by real-time PCR. Total RNA was extracted from the heart, liver, spleen, kidney and skin tissues of the Tibetan sheep. The results showed that the coding region of Tibetan sheep KAP3.2 gene was 297 bp in length, encoding 98 amino acids. The homologies of nucleotide sequences of the coding region of KAP3.2 gene between Tibetan sheep and sheep, goat, chiru, prairie vole, mouse and human were 99%, 97%, 96%, 79%, 75% and 73%, respectively. The KAP3.2 gene was highly expressed in the skin tissue and relatively was low in other tissues studied. The expression level of KAP3.2 m RNA is tissue specific.