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, VP1, is of great interest for the study on capsid assembly in vitro. Production of soluble His6-VP1 in Escherichia coli strains deficient in the GroELS chaperone machine was substantially higher than in the wild-type strain. The His6-VP1 produced in a groEL 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 microscopy. We propose the use of groE mutants for the production of the major capsid protein of SV40 and perhaps also other papovaviruses.
Wróbel
B
B
Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School, Ein Kerem, POB 12272, 91120, Jerusalem, Israel. wrobel@gene.md.huji.ac.il
Yosef
Y
Y
Oppenheim
A B
AB
Oppenheim
A
A
D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D036022:Capsid Proteins; D018835:Chaperonin 10 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics*; D014771:Virion / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; C062481:VP1 protein, polyomavirus