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。
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, coil 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.
出处
《蚕业科学》
CAS
CSCD
北大核心
2006年第1期36-41,共6页
ACTA SERICOLOGICA SINICA
基金
国家自然科学基金项目(编号50473014)
苏州市农业科技计划项目(编号SNZ0410)
关键词
仿丝素结晶区
载体构建
大肠杆菌表达
Fibroin crystalline domain like protein
Vector construction
Gene expression in E. coil
