人甲状旁腺素N端34肽基因的合成,克隆及融合表达

Synthesis Cloning and Fusion Expression of the Human Parathyroid Hormone N terminus 34 Peptide Gene

摘要: 选用大肠杆菌偏爱的密码子,用计算机辅助设计、合成长度分别为59 、59 及60 碱基的三段寡核苷酸。以中段寡核苷酸为模板,两端寡核苷酸为引物,经PCR 合成完整的hPTH(134) 基因,并克隆到pUC18 载体中。对克隆基因进行序列分析,结果与设计的完全一致。将该基因与表达载体pKA 连接构建表达质粒pKAT,转化大肠杆菌JM105 细胞,SDSPAGE 表明hPTH(134) 融合蛋白获得了表达,并且酸水解后融合蛋白被裂解为大小两个片段。

Abstract: The human parathyroid hormone (PTH) N terminal 34 peptide gene has been synthesized and cloned. Computer progamming was applied to the design of optimized segments for the synthesis of the hPTH(1 34) gene using preferred codons for expression in E.coli