摘要
目的研究不同因素对溴化氰切割融合蛋白的影响。方法将含有pThioHisA-G13重组质粒的BL21工程菌,经IPTG诱导后,超声破碎,分离并溶解包涵体,再选择不同量的溴化氰、酸浓度及不同溶剂等条件进行溴化氰切割试验,Tricine-SDS-PAGE检测切割效果。结果溴化氰的量,酸浓度及不同溶剂对溴化氰切割包涵体的效果均有影响。结论高效率的切割需要在酸性条件下进行;以尿素作为促溶剂比70%的甲酸更有利于小肽的切割。
Purpose To study the effect of different factors on the efficiency of cleaving fusion protein with cyanogen bromide （CNBr）. Methods The engineered E. coli BL21 （DE3） cells harboring recombinant expression vector,pThioHisA-G13 ,were induced by IPTG, and then were lysed by sonication to extract inclusion body. Different concentrations of CNBr and acids, and different kinds of solvents were tested for the CNBr cleavage reaction. Tricine-SDS-PAGE was used to analyze the cleavage results. Results The concentrations of CNBr or acids and solvent types had the effect on cutting efficiency. Conclusion A high cutting efficiency required the acid environment and urea as a solutizer had an advantage over 70 % formic acid in facilitating cleavage.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2009年第2期93-95,共3页
Chinese Journal of Biochemical Pharmaceutics
基金
安徽省高校省级自然科学研究重点项目(No.KJ2007A091)
安徽大学211工程学术创新团队资助(No.02203109)
关键词
溴化氰
融合蛋白
切割
cyanogen bromide
fusion protein
cleavage
