Novel human cytomegalovirus DNA sequences

Abstract

Provided are novel Toledo and Towne human cytomegalovirus DNA sequences (HCMV) and proteins encoded thereby. The sequences are useful in methods and compositions for detecting HCMV infections and in immunogenic compositions for preventing HCMV infections.

What is claimed is:  
     
         1 . An isolated DNA sequence selected from the group consisting of: 
 a. the DNA corresponding to the nucleotide sequence of FIG. 1 (SEQ ID NO:6);    b. the DNA corresponding to the nucleotide sequence of FIG. 2 (SEQ ID NO:1);    c. a DNA corresponding to the nucleotide sequence of FIG. 1 (SEQ ID NO:6) in inverted orientation; and    d. a DNA capable of hybridizing under stringent conditions, or which would be capable of hybridizing under said conditions but for the degeneracy of the genetic code, to a sequence corresponding to the sequence of FIG. 1 (SEQ ID NO:6), or FIG. 2 (SEQ ID NO:1).    
     
     
         2 . An RNA molecule transcribed from a DNA sequence of  claim 1 .  
     
     
         3 . A vector comprising a DNA sequence of  claim 1 .  
     
     
         4 . A host cell transformed with a DNA sequence of  claim 1  in operative association with an expression control sequence capable of directing replication and expression of said DNA sequence.  
     
     
         5 . A HCMV protein encoded by a DNA sequence of  claim 1 .  
     
     
         6 . A method of producing a HCMV protein comprising culturing a host cell of  claim 4  in a suitable culture medium and isolating said protein from said medium.  
     
     
         7 . A method of producing a recombinant HCMV comprising: 
 a. transfecting a plasmid containing a DNA sequence according to  claim 1  and (i) transfecting viral DNA comprising AD169 or Towne strain HCMV or (ii) superinfecting AD169 or Towne strain HCMV into a cell line permissive for growth of CMV; and    b. isolating said recombinant HCMV from said cell line.    
     
     
         8 . An immunogenic composition comprising a recombinant HCMV made by the method of  claim 7 .  
     
     
         9 . A method of prophylactic treatment of a HCMV-related disease or condition comprising administering to a patient an immunogenic composition of  claim 8  in an amount sufficient to stimulate an immune response in said patient.  
     
     
         10 . A method for detecting an HCMV infection in a human comprising: 
 (i) combining under predetermined stringency conditions a clinical sample suspected of containing HCMV DNA with at least one single-stranded DNA fragment of a DNA sequence according to  claim 1  having at least 10 bases and being non-cross-hybridizing with human DNA; and    (ii) detecting duplex formation between said single-stranded DNA fragment and the sample DNA.    
     
     
         11 . A pharmaceutical composition comprising a protein encoded by a DNA sequence of  claim 1  in admixture with a pharmaceutically acceptable carrier.  
     
     
         12 . A method of prophylactic treatment of a HCMV-related disease or condition comprising administering to a patient a pharmaceutic composition of  claim 11  in an amount to stimulate an immune response in said patient.