【摘要】 为探讨蚕丝结构与性能的关系,设计了几种类似蚕丝丝素结晶区高度重复序列结构(GAGAGX)n的基因序列,并构建了大肠杆菌表达载体,在BL-21菌株中成功诱导表达了4种约由110个氨基酸组成的小分子蛋白。对表达产物进行W estern印迹分析和ELISA检测的结果显示,(GAGAGA)n、(GAGAGY)n、(GAGAGV)n3种蛋白都有小于14.4 kD的大小相同的条带,而(GAGAGS)n有一条稍大于20.1 kD的条带。以超声波破菌的缓冲液作空白对照,4种蛋白的ELISA的结果均比对照的BL-21(无质粒)高,显示为阳性。更多还原

【Abstract】 We have designed four gene fragments encoding silkworm fibroin crystalline domain like proteins,（GAGAGX）_n,which are highly repetitive in silk.These fragments were cloned into pCDNA3 expression vectors and successfully expressed in E.coli BL-21 under inducement of IPTG.The vectors were characterized by enzyme digestion and gene sequencing.Western blot analysis showed that three expressed proteins have molecular masses about 14.4 kD,another is a bit larger than 20.1 kD.ELISA analysis showed that all these four proteins extracted by ultrasonic were positive.更多还原