| 家蚕丝素结晶区高度重复多肽[GAGAGA]_(16)和[GAGAGS]_(16)的融合蛋白优化表达 |
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| 引用本文： | 王建南,白伦.家蚕丝素结晶区高度重复多肽[GAGAGA]_(16)和[GAGAGS]_(16)的融合蛋白优化表达[J].蚕业科学,2009,35(3). |
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| 作者姓名： | 王建南 白伦 |
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| 作者单位： | 现代丝绸国家工程实验室,江苏省丝绸重点实验室,苏州大学纺织与服装工程学院,江苏苏州,215021 |
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| 基金项目： | 江苏省自然科学基金项目，苏州市科技发展计划项目 |
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| 摘 要： | 为了给深入研究家蚕丝素结晶区的典型肽段结构提供素材,对设计的GST-tag家蚕丝素结晶区典型多肽GAGAGA]16和GAGAGS]16的融合蛋白KGA、KGS进行了表达条件优化试验。结果表明,pGEX-KGA和pGEX-KGS表达载体能够有效表达家蚕丝素结晶区的2种典型多肽,KGA和KGS的最佳表达诱导剂IPTG的浓度分别为1.4mmol/L、1.0 mmol/L,诱导时间为5 h。在此优化条件下,KGA和KGS在大肠杆菌(E.coli)中的表达量分别达75 mg/L、110 mg/L。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。
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| 关 键 词： | 家蚕 丝素结晶区 典型肽段 大肠杆菌 GST亲和层析 表达效率 |
Optimization of Expression of Highly Repetitive Polypeptides [GAGAGA]_(16) and [GAGAGS]_(16) From Bombyx mori Silk Fibroin Crystalline Domain |
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| WANG Jian-Nan,BAI Lun.Optimization of Expression of Highly Repetitive Polypeptides [GAGAGA]_(16) and [GAGAGS]_(16) From Bombyx mori Silk Fibroin Crystalline Domain[J].Acta Sericologica Sinica,2009,35(3). |
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| Authors: | WANG Jian-Nan BAI Lun |
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| Institution: | WANG Jian-Nan BAI Lun (National Engineering Laboratory for Modern Silk,Key Laboratory for Silk,Jiangsu Province,College of Textile , Clothing Engineering,Soochow University,Suzhou Jiangsu 215021,China) |
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| Abstract: | In order to study structures of the typical peptides from silk fibroin crystalline domain by gene engineering,the expression conditions were optimized for expressing designed GST-tagged crystalline peptidesGAGAGA]16 andGAGAGS] 16 fusion proteins(KGA and KGS).The results showed that:fusion proteins KGA and KGS could be expressed efficiently using the expression vectors pGEX-KGA and pGEX-KGS.The best IPTG concentrations for KGA and KGS were 1.4 mmol/L and 1.0 mmol/L respectively,and the best induction time ... |
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| Keywords: | Bombyx mori Fibroin crystalline domain Typical polypeptide Escherichia coli GST affinity chromatography Expression efficiency |
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