家蚕丝素结晶区典型多肽[GAGAGY]16和[GAGAGV]16的融合蛋白优化表达  

Optimized Expression of Fusion Proteins with Typical Polypeptides [GAGAGY]_(16) and [GAGAGV]_(16) from Bombyx mori Silk Fibroin Crystalline Domain

作　　者：王建南[1] 白伦[1] 李明忠[1] 

机构地区：[1]现代丝绸国家工程实验室,苏州市生物组织工程材料与技术重点实验室,苏州大学纺织与服装工程学院,江苏苏州215021

出　　处：《蚕业科学》2010年第1期175-180,共6页ACTA SERICOLOGICA SINICA

基　　金：江苏省自然科学基金项目(No.BK2009147);江苏省高校自然科学基础研究项目(No.07KJA43010);苏州市科技发展计划项目(No.SS0829)

摘　　要：家蚕丝素结晶区典型多肽[GAGAGY]16、[GAGAGV]16的GST融合蛋白KGY和KGV是深入研究家蚕丝素结晶区典型多肽聚集态结构的素材。对这2种融合蛋白在大肠杆菌中的表达条件进行优化试验的结果表明,表达载体pGEX-KGY和pGEX-KGV能够有效表达家蚕丝素结晶区的这2种典型多肽。KGY和KGV的最佳诱导表达条件:IPTG浓度分别为0.2 mmol/L和1.0 mmol/L,诱导时间分别为5 h和4 h。在此条件下,2种融合蛋白在大肠杆菌中的表达量分别达115 mg/L和90 mg/L左右。诱导起始时大肠杆菌的菌密度(OD600为0.4～3.0)对2种融合蛋白的表达效率没有显著影响。GST-tagged fusion proteins KGY and KGV that have typical polypeptides[GAGAGY]16and[GAGAGV]16 from silk fibroin crystalline domain are good materials for in-depth study on clustered structures of the typical polypeptides from silk fibroin crystalline domain.Optimization tests for expression of the two fusion proteins in E.coli revealed that fusion proteins KGY and KGV could be expressed efficiently on expression vectors pGEX-KGY and pGEX-KGV.The optimal expression conditions of KGY and KGV were concentration of IPTG at 0.2 mmol/L and 1.0 mmol/L,and the induction time at 5 h and 4 h respectively.Under these conditions,the expressed quantity of KGY and KGV reached 115 mg/L and 90 mg/L,respectively.It was also found that the initial density of E.coli culture（OD600 ranging from 0.4 to 3.0） before the induction had no obvious influence on the expression of these two fusion proteins.

关 键 词：家蚕 丝素结晶区 典型肽段 表达载体 诱导剂浓度 诱导时间 表达效率 

分 类 号：S881.24[农业科学—特种经济动物饲养]