Heterologous expression of neisserial proteins

Abstract

Alternative and improved approaches to the heterologous expression of the proteins of Neisseria meningitidis and Neisseria gonorrhoeae are disclosed. These approaches typically affect the level of expression, the ease of purification, the cellular localization, and/or the immunological properties of the expressed protein.

1 - 13 . (canceled) 
     
     
         14 . A method for the heterologous expression of a protein of the invention, in which (a) no fusion partner is used, and (b) the protein's native leader peptide (if present) is used. 
     
     
         15 . The method of  claim 14 , in which the protein of the invention is selected from the group consisting of: 111, 149, 206, 225-1, 235, 247-1, 274, 283, 286, 292, 401, 406, 502-1, 503, 519-1, 525-1, 552, 556, 557, 570, 576-1, 580, 583, 664, 759, 907, 913, 920-1, 936-1, 953, 961, 983, 989, Orf4, Orf7-1, Orf9-1, Orf23, Orf25, Orf37, Orf38, Orf40, Orf40.1, Orf40.2, Orf72-1, Orf76-1, Orf85-2, Orf91, Orf97-1, Orf119, Orf143.1, NMB0109, NMB2050, 008, 105, 117-1, 121-1, 122-1, 128-1, 148, 216, 243, 308, 593, 652, 726, 926, 982, Orf83-1 and Orf143-1. 
     
     
         16 . A method for the heterologous expression of a protein of the invention, in which (a) the protein's leader peptide is replaced by the leader peptide from a different protein and, optionally, (b) no fusion partner is used. 
     
     
         17 . The method of  claim 16 , in which the different protein is 961, ORF4,  E. coli  OmpA, or  E. carotovora  PelB, or in which the leader peptide is MKKYLFSAA. 
     
     
         18 - 49 . (canceled) 
     
     
         50 . The method of  claim 16 , in which the heterologous expression is in an  E. coli  host. 
     
     
         51 - 52 . (canceled) 
     
     
         53 . An immunogenic composition comprising an alum adjuvant and  E. coli  lipidated protein produced by the method of  claim 50 . 
     
     
         54 . A method of inducing an immune response to an  E. coli  lipidated protein produced by the method of  claim 50  in a subject comprising administering to the subject an immunogenic composition comprising an alum adjuvant and the  E. coli  lipidated protein. 
     
     
         55 . A purified polypeptide comprising: (a) a fragment of the protein produced by the method of  claim 16 , or (b) an amino acid sequence having at least 90% sequence identity to the protein produced by the method of  claim 16 .