IEF纯化重组A型肉毒毒素保护性抗原.PDF

6 2 Vol.6 No.2 2002 6 Life Science Research June 2002 IEF A ¹ 王 慧, 荫 俊 ( 100071) :在大肠杆菌中高效表达的重组A 型肉毒毒素保护性抗原(rBoNTa 468) , 是以包涵体形式存在.将表达 菌 发酵后,裂解菌体, 制备包涵体, 溶解后的包涵体溶液经样品处理,通过等电聚焦制备型电泳纯化. 纯化的 重组A 型肉毒毒素保护性抗原(rBoNTa 468) 纯度高于90%, 产量及回收率高. 纯化的重组表达产物酶联检测 具有结合活性,这为下 一步A 型肉毒毒素抗毒素的研制打下基础. :A 型肉毒毒素; 保护性抗原; 等电聚焦; 纯化 :Q751 :A : 1007-7847(2002)02-0133-04 Purification of Recombinant Protective Fragment of Botulinum Neurotoxin Serotype A by IEF WANG ui, YIN Jun ( Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beji ing 100071, China) Abstract: Recombinant protective fragment(r BoNTa c468) of botulinum neurotoxin serotype A( BoNTa) was ex- pressed at high level in insoluble form in E. coli.Therecombinant proteins were purifiedwith following two steps: Firstly, lystated E . coli cell pellets after ferment of E. coli strain containing recombinant expression plasmid pBV- BoNTa c468, and made inclusion body. Secondly, resolved inclusion body with 8mol/L deionized urea. Purified protein finally through Isoelectronic Fucos (IEF) preparation electrophoresis. As a result, pIof protein is about p 8. 0~ 8.5.Purificativity of proteins is up to over 90%. Purified protien is positivein ELISA. Thesuccessful purifi- cation and expression of protective fragment(rBoNTa c468) of botulinum neurotoxin serotype A is a key to further study on gene engineering antitoxin and subunit vaccine. Key wor s: botulinum neurotoxin serotypeA (BoNTa) ; protective antigen; IsoelectronicFucos(IEF) ; purification ( fi e Science Research, 2002, 6(2) : 133~ 136) , . , , ,, , ,