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高盐胁迫渗透对SBD2重组蛋白在大肠杆菌中可溶性表达的影响

Effect of Hyperosmotic Stress on Expression of Soluble SBD2 Recombinant Protien in Escherichia coli
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摘要 在基因工程的操作中,包涵体的形成是人们利用大肠杆菌表达外源蛋白质的一大难点.本研究将环状芽孢杆菌(Bacillus circulans)环状糊精糖基转移酶(CGTase)的淀粉粒结合域(SBD)基因编码序列的两个拷贝通过一连接肽连接(SBD2)后,克隆到大肠杆菌表达载体pTrcHis B上,得到的pTrcHis B/SBD2质粒转化大肠杆菌Top10,研究了不同培养基、不同诱导温度和时间对SBD2蛋白表达的影响.结果表明,利用相容性溶质山梨醇和甜菜碱等,在高盐胁迫下,实现了SBD2重组蛋白质在大肠杆菌中的可溶性表达,这一结果为在体外研究SBD2蛋白的功能奠定了基础. An artificial tandm repeat of a starch-binding domain（SBD2） was engineered by two copies of the SBD derived from Bacillus circulans cyclodextrin glycosyltransferase via a linker peptide.The SBD2 gene was inserted into an E.coli expression vector,pTrcHis B,to obtain the pTrcHis B/SBD2 vector.Recombinant E.coli Top 10 cells,containing the pTrcHis B/SBD2construct,were grown in LB with 50μg/mL ampicillin and LB with 50μg/mL ampicillin,1mol/L sorbitol and 2.5mmol/L glycyl betaine,respectively.Expression of SBD2 protein was induced by adding of IPTG and analyzed by SDS-PAGE.The results show that large amounts of soluble protein were obtained by growing and inducing the cells under osmotic stress in the presence of sorbitol and glycyl betaine.
作者 季勤 张云峰 罗玉明 徐春 黄翠香
机构地区 淮阴师范学院生命科学学院 南京师范大学生命科学学院
出处 《南京师大学报（自然科学版）》 CAS CSCD 北大核心 2010年第3期70-75,共6页 Journal of Nanjing Normal University(Natural Science Edition)
基金 国家自然科学基金(30771367) 江苏省高校高新技术产业发展基金(JHB04-043) 淮阴师范学院教授基金(06HSJS025) “青蓝工程”
关键词 SBD重组蛋白 相容性溶质 高渗胁迫 可溶性表达 SBD2 protein compatible solute hyperosmotic stress soluble expression
分类号 Q786 [生物学—分子生物学]
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参考文献14

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南京师大学报（自然科学版）
2010年 第3期

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