AiiA融合蛋白包涵体变性和复性的研究  

Study on Inclusion Body's Denaturation and Renaturation of AiiA Fusion Protein

作　　者：杨梅[1] 郭丽清[1] 关夏玉[1] 张峰[2] 林彬辉[1] 陈新华[1] 黄志鹏[1] 

机构地区：[1]福建农林大学生物农药与化学生物学教育部重点实验室,福建福州350002 [2]福建师范大学生命科学学院,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》2008年第4期76-79,共4页Journal of Fujian Normal University：Natural Science Edition

基　　金：福建省科技厅基金资助项目(2006J0051)

摘　　要：通过IPTG诱导含有pGEXaiiA-B15质粒的工程菌使其大量表达AiiA融合蛋白.由于所表达的融合蛋白多为不可溶的包涵体,须经分离、变性溶解,再经过一个合适的复性过程才能实现变性蛋白的正确折叠,得到具有生物活性的蛋白.通过含N-十二烷基肌氨酸钠的缓冲液A溶液及尿素等变性剂使包涵体溶解,磷酸盐缓冲液透析复性.SDS-PAGE电泳表明包涵体已由不可溶转化成可溶的蛋白.抑菌试验证明复性后的AiiA蛋白对胡萝卜欧文氏软腐病菌引起的马铃薯软腐病有较明显的抑制作用.The recombinant expression vectors pGEXaiiA-B15 were used to express the Alia fusion protein in Escherichia coli BL21 after induction with IPTG. Due to the expressed fusion protein are the insoluble inclusion body, it must be seperated, denaturated and refolded correctly after a proper renaturation in order to obtain the protein with biological activity. The insoluble inclusion body was denatured with Urea and N-Lauroyl Sarcosine Sodium and then was renatured after the dialysis with the phosphate buffer solution. SDS-PAGE indicated that the inclusion body after the above treatment had turned to soluble protein. By bacteriostatic experiment, the result showed that AiiA protein after renaturation had a significant inhibiting ability to patato soft rot caused by the Erwinia carotovora.

关 键 词：苏云金芽孢杆菌 AiiA蛋白 变性 复性 

分 类 号：Q932[生物学—微生物学]