大肠杆菌表达GST-Aβ_(42)融合蛋白的纯化条件及鉴定  

Optimal Purification Condition and Identification of GST-Aβ_(42)Expression in Escherichia Coli

作　　者：宋翠翠 曹国宪 张莉[2] 俞惠新[2] 林秀峰[2] 

机构地区：[1]江南大学医药学院,江苏无锡214122 [2]江苏省原子医学研究所卫生部核医学重点实验室,江苏无锡214063

出　　处：《现代生物医学进展》2008年第12期2453-2456,共4页Progress in Modern Biomedicine

基　　金：江苏省自然科学基金(NO.BK2006026);无锡市自然科学基金(NO.CK040004)资助

摘　　要：目的:为了提高β-淀粉样蛋白(β-amyloid peptide,Aβ_(42))基因在大肠杆菌中的表达,为深入研究Aβ的作用机制及其疫苗研究奠定基础。方法:大肠杆菌在37℃培养4 h后,以终浓度为1 mmol/L的IPTG在25℃下继续诱导培养2 h,促进GST-Aβ42融合蛋白的可溶性表达。表达产物以SDS-PAGE、Western bloting鉴定。结果:SDS-PAGE表明融合蛋白分子量约为32kD,与预计的一致;Western Blotting进一步分析表明它能与抗Aβ42和抗GST抗体特异反应。结论:GST-Aβ_(42)基因的优化表达为研究Aβ42的作用机理打下了基础,同时也为Aβ42疫苗的研究提供了充分的实验条件。Objective： To improve the expression of β-amyloid peptide in Escherichia coli, and also to lay a foundation for further study on its Mechanism and vaccine function. Method： The results indicated that the soluble expression level of GST-Aβ42 fusion protein could be promoted when cells were cultured for 4 h at 37 ℃, and then shifted to 25 ℃ for 2 h to induce the expression of GST-Aβ42 with 1 mmol/L IPTG as final concentration. The expression product was determined by SDS-PAGE and Westem Blotting analysis. Result： SDS-PAGE analysis result showed that the molecular mass of GST-Aβ42 fusion protein was about 32 kD as expected, and Western Blot- ting analysis indicated that both anti-Aβ42 and anti-GST antibody could react specifically against the fusion protein. Conclusion： The optimized expression of GST-Aβ42 will be useful for future research on the mechanism of Aβ42 on Alzheimer＇s Disease and provide sufficient foundation for the further study on Aβ42 vaccine.

关 键 词：Β-淀粉样蛋白 融合蛋白 可溶性表达 

分 类 号：Q786[生物学—分子生物学]