Dna sequence, and recombinant preparation of group 4 major allergens from cereals

Abstract

The present invention relates to the provision of DNA sequences of group 4 mayor allergens from cereals. The invention also encompasses fragments, new combinations of partial sequences and point mutants having a hypoallergenic action. The recombinant DNA molecules and the derived polypeptides, fragments, new combinations of partial sequences and variants can be utilised for the therapy of pollen-allergic diseases. The proteins prepared by recombinant methods can be employed for in vitro and in vivo diagnosis of pollen allergies.

1 .- 18 . (canceled) 
     
     
         19 . A DNA molecule comprising
 (a) a nucleic acid sequence which hybridizes with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7 and SEQ ID NO: 9 under stringent hybridization conditions, or   (b) a nucleic acid which is the complement of (a), which encodes a polypeptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10,   
       wherein the nucleic acid sequence of (b) has been codon-optimized for the expression of the polypeptide product thereof in a host cell that is not a cell of  Secale cereale, Hordeum vulgare  or  Triticum aestivum.    
     
     
         20 . A DNA molecule comprising at least one partial sequence of a nucleic acid sequence encoding a polypeptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10, wherein said DNA molecule encodes an immunomodulatory, T-cell reactive fragment of a group 4 allergen from Poaceae. 
     
     
         21 . The isolated DNA molecule of  claim 19 , wherein the DNA molecule encodes a polypeptide having at least one amino acid difference from a polypeptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10. 
     
     
         22 . The isolated DNA molecule of  claim 21 , wherein at least one amino acid difference is a deletion or a substitution of at least one cysteine. 
     
     
         23 . The isolated DNA molecule of  claim 21 , comprising a nucleic acid sequence encoding a polypeptide selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15. 
     
     
         24 . A recombinant DNA expression vector or a cloning system comprising an isolated DNA molecule of  claim 19 . 
     
     
         25 . A culture comprising a host organism transformed with the recombinant DNA expression vector or cloning system of  claim 24 . 
     
     
         26 . A process for the preparation of a polypeptide encoded by the isolated DNA molecule of  claim 19 , comprising
 cultivating a host organism transformed with the DNA molecule and   isolating the polypeptide from the culture.   
     
     
         27 . A pharmaceutical composition comprising at least one isolated DNA molecule of  claim 19  and a pharmaceutically acceptable adjuvant. 
     
     
         28 . A method for the prevention and/or treatment of an allergy triggered by an allergen from Poaceae species, comprising administering to a subject in need thereof an effective amount at least one DNA molecule of  claim 19 . 
     
     
         29 . A method for the prevention and/or treatment of an allergy triggered by an allergen from Poaceae species, comprising administering to a subject in need thereof an effective amount at least recombinant DNA expression vector of  claim 24 . 
     
     
         30 . A method for the prevention and/or treatment of an allergy triggered by an allergen from Poaceae species, comprising administering to a subject in need thereof an effective amount at least one pharmaceutical composition of  claim 27 .