US2007161083A1PendingUtilityA1

Codon Optimized Recombinant Dermaphagoides Allergens

Assignee: GLAXOSMITHKLINE BIOLOG SAPriority: Jun 10, 2000Filed: Dec 12, 2006Published: Jul 12, 2007
Est. expiryJun 10, 2020(expired)· nominal 20-yr term from priority
A61P 37/08A61K 38/00A61P 11/06C07K 14/43531
50
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Claims

Abstract

The present invention relates to codon optimised polynucleotides which are efficiently expressed in mammalian cells and encode insect proteins from Dermaphagoides dust mite. In particular, the optimised codon polynucleotides encode a protein from Dermaphagoides pteronyssinus , such as DerP1 or proDerP1. The present invention also provides methods of preparing pharmaceutical compositions comprising the expression of the codon optimised polynucleotides, and vectors and transformed host cells comprising them.

Claims

exact text as granted — not AI-modified
1 . An isolated polynucleotide sequence which encodes a  Dermatophagoides  mite protein, wherein the codon usage pattern of the polynucleotide sequence resembles that of highly expressed mammalian genes.  
     
     
         2 . An isolated polynucleic acid sequence as claimed in  claim 1 , wherein all of the amino acid types present in the protein are optimised, such that the codons used to encode that amino acid are used in the same frequency as the known mammalian frequency for that amino acid.  
     
     
         3 . An isolated polynucleic acid sequence as claimed in  claim 1 , wherein the yield of protein encoded by the polynucleic acid when expressed by an expression system is greater than 20% higher than the amount of protein produced from the same expression system using a non-optimised native gene for that  Dermatophagoides  protein.  
     
     
         4 . An isolated polynucleic acid sequence encoding a  Dermaphagoides  protein, characterised in that the codons present in said polynucleotide which are used to encode each amino acid are selected to appear as set forth in the following table  
       
         
           
                 
                 
                 
               
                     
                 
                     
                 
                   Amino 
                     
                   Frequency 
                 
                   Acid 
                   Codon 
                   (percentage used) 
                 
                     
                 
                     
                 
                 
                 
                 
               
                   Ala 
                   GCG 
                   17 
                 
                     
                   GCA 
                   13 
                 
                     
                   GCT 
                   17 
                 
                     
                   GCC 
                   53 
                 
                   Arg 
                   AGG 
                   18 
                 
                     
                   AGA 
                   10 
                 
                     
                   CGG 
                   21 
                 
                     
                   CGA 
                   6 
                 
                     
                   CGT 
                   7 
                 
                     
                   CGC 
                   37 
                 
                   Asn 
                   AAT 
                   22 
                 
                     
                   AAC 
                   78 
                 
                   Asp 
                   GAT 
                   25 
                 
                     
                   GAC 
                   75 
                 
                   Cys 
                   TGT 
                   32 
                 
                     
                   TGC 
                   68 
                 
                   Gln 
                   CAG 
                   88 
                 
                     
                   CAA 
                   12 
                 
                   Glu 
                   GAG 
                   75 
                 
                     
                   GAA 
                   25 
                 
                   Gly 
                   GGG 
                   24 
                 
                     
                   GGA 
                   14 
                 
                     
                   GGT 
                   12 
                 
                     
                   GGC 
                   50 
                 
                   His 
                   CAT 
                   21 
                 
                     
                   CAC 
                   79 
                 
                   Ile 
                   ATA 
                   5 
                 
                     
                   ATT 
                   18 
                 
                     
                   ATC 
                   77 
                 
                   Leu 
                   TTG 
                   6 
                 
                     
                   TTA 
                   2 
                 
                     
                   CTG 
                   58 
                 
                     
                   CTA 
                   3 
                 
                     
                   CTT 
                   5 
                 
                     
                   CTC 
                   26 
                 
                   Lys 
                   AAG 
                   82 
                 
                     
                   AAA 
                   18 
                 
                   Phe 
                   TTT 
                   20 
                 
                     
                   TTC 
                   80 
                 
                   Pro 
                   CCG 
                   17 
                 
                     
                   CCA 
                   16 
                 
                     
                   CCT 
                   19 
                 
                     
                   CCC 
                   48 
                 
                   Ser 
                   AGT 
                   10 
                 
                     
                   AGC 
                   34 
                 
                     
                   TCG 
                   9 
                 
                     
                   TCA 
                   5 
                 
                     
                   TCT 
                   13 
                 
                     
                   TCC 
                   28 
                 
                   Thr 
                   ACG 
                   15 
                 
                     
                   ACA 
                   14 
                 
                     
                   ACT 
                   14 
                 
                     
                   ACC 
                   57 
                 
                   Tyr 
                   TAT 
                   26 
                 
                     
                   TAC 
                   74 
                 
                   Val 
                   GTG 
                   64 
                 
                     
                   GTA 
                   5 
                 
                     
                   GTT 
                   7 
                 
                     
                   GTC 
                   25 
                 
                     
                 
                     
                 
             
                
                
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . An isolated polynucleotide comprising SEQ ID NO: 15.  
     
     
         6 . An isolated vector comprising the isolated polynucleotide of  claim 5 .  
     
     
         7 . An isolated host cell comprising the vector of  claim 6.

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