【摘要】 青岛海葵中存在两种具有增强心肌收缩功能的多肽类毒素 (分别命名为Ap QD1和Ap QD2 ) .通过分析已经得到Ap QD2的氨基酸序列 ,按照毕氏酵母的偏爱密码子设计并合成了Ap QD2的cDNA .将合成cDNA序列通过PCR扩增及一系列分子克隆操作导入毕氏酵母表达载体 pPICZαA中 ,以电穿孔方法转化毕氏酵母GS115和KM71,并进行转化子的表型及高拷贝化筛选 .其中的KM 71(Muts)的转化子经摇瓶发酵 ,每升发酵液可表达约 2 0mg的重组Ap QD2产物 ,通过纯化可得到 7mg纯的有天然生物学活性的Ap QD2基因工程产物 .更多还原

【Abstract】 Venom of sea anemone Anthopleura qingdaoensis contains at least two cardiotonic polypeptides capable of enhancing cardiac contraction(a positive inotropic effect). An artificial gene encoding anthopleurin QD2 (Ap QD2), which consists of 49 anino acid residues, was cloned and transferred into pPICZαA, a secretory expression vector for Pichia pastoris. The construct was linearized and was integrated into the yeast chromosome by electroporation under the selection of zeocin. Approximately 20 mg/L of biologically active Ap QD2 was produced from one of the KM71(Mut s) transformants, and about 7 mg of pure Ap QD2 was obtained after chromatography purification.更多还原