A simple procedure for expression and purification of selected non-structural (alpha and beta) herpes simplex virus 1 (HSV-1) proteins

The expression and isolation of herpes simplex virus 1 (HSV-1) immediate ea rly (alpha) IE63 (ICP27) and of the early (beta) thymidine kinase (Tk) poly peptides in Escherichia coli JM 109 cells transformed with the PinPoint Xa- 1 (Promega) plasmid construct carrying either the HSV-1 UL54 or UL23 genes are described. The resulting biotinylated fusion protein(s) could be easily induced and were purified in appropriate amounts by means of a monomeric a vidin-conjugated resin (SoftLink(TM) Soft Release Avidin Resin, Promega) pr ovided that: (1) the exponential growth of the selected transformed cells w as monitored carefully; (2) the post-induction harvest interval was properl y chosen; and (3) the period for adsorption to the avidin resin suitably ad justed. The isolated protein(s), although partially digested in the case of the IE63 polypeptide. were suitable antigen(s) for immunization of various animal species. Co-purification of trace amounts of endogenous biotinylate d protein(s) produced in E. coli was eliminated by shortening the duration of adsorption to the avidin resin. (C) 2001 Elsevier Science B.V. All right s reserved.