An inducible expression system permitting the efficient purification of a recombinant antigen from Mycobacterium smegmatis
An inducible expression system permitting the efficient purification of a recombinant antigen from Mycobacterium smegmatis
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DOI：
10.1111/j.1574-6968.1998.tb13221.x
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发表时间：
1998-10-15
影响因子：
2.1
通讯作者：
Gicquel, B
中科院分区：
文献类型：
--
作者：
Triccas, JA;Parish, T;Gicquel, B
构建了一种利用耻垢分枝杆菌高度诱导型乙酰胺酶启动子的新型表达载体。在存在乙酰胺酶诱导剂乙酰胺的情况下生长的重组耻垢分枝杆菌中证实了模型抗原（麻风分枝杆菌 35 kDa 蛋白）的高水平诱导。借助C端6-组氨酸标签，可以简单有效地从细菌超声处理中纯化重组蛋白，证明这种纯化策略可用于分枝杆菌。组氨酸标签对蛋白质构象或免疫原性没有明显影响，这表明所描述的载体可能可用于从快速生长的分枝杆菌宿主中纯化类似天然的重组分枝杆菌蛋白质。 (C) 1998 年欧洲微生物学会联合会。由 Elsevier Science B.V. 出版。保留所有权利。
A novel expression vector utilising the highly inducible acetamidase promoter of Mycobacterium smegmatis was constructed. High-level induction of a model antigen, the Mycobacterium leprae 35 kDa protein, was demonstrated in recombinant M. smegmatis grown in the presence of the acetamidase inducer acetamide. The recombinant protein could be simply and efficiently purified from the bacterial sonicate by virtue of a C-terminal 6-histidine tag, demonstrating that this purification strategy can be used for the mycobacteria. The histidine tag had no apparent effect on the protein conformation or immunogenicity, suggesting that the vector described may prove useful for the purification of native-like recombinant mycobacterial proteins from fast-growing mycobacterial hosts. (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.