基因定点诱变删除及天然α型心钠素的分泌型表达

Directed Deletion of the Cloned α-ANF Gene and Its Secretive Expression in E. coli

用基因定点诱变技术,删除了pO_1α ANF表达质粒中的33对碱基,使人α型心钠素结构基因直接与大肠杆菌分泌型表达质粒pIN-Ⅲ-OmPA中的信号肽酶切位点编码区相连,构成天然人α型心钠素的表达质粒pANF,在IPTG诱导下表达28肽的天然人α型心钠素。纯化后的表达产物具有天然心钠素的放免活性和很强的舒张血管的生物活性。

A modified method of oligonucleotide-directed site-specific matagensis was used to delete a 33 base fragment from a recombinant plasmid pO1 αANF, resulting in the secretion of authentic α-ANF from expression strain pANF/JA221.The gene product was purified and further characterized for biological activites.