Production and purification of SV40 major capsid protein (VP1) in Escherichia coli strains deficient for the GroELS chaperone machine

Production of the major capsid protein of SV40, VPI, is of great interest f or the study on capsid assembly in vitro. Production of soluble His(6)-VP1 in Escherichia coli strains deficient in the GroELS chaperone machine was s ubstantially higher than in the wild-type strain. The His(6)-VP1 produced i n a gPoEL mutant strain was readily purified. The protein was able to form higher-order structures as evidenced by analysis of the soluble fraction by gel filtration, by sedimentation in sucrose gradient, and by electron micr oscopy. We propose the use of groE mutants for the production of the major capsid protein of SV40 and perhaps also other papovaviruses. (C) 2000 Elsev ier Science B.V. All rights reserved.