Recombinant vector containing infectious human cytomegalovirus genome with preserved wild-type characteristics of clinical isolates

Abstract

A recombinant vector containing infectious genome of human cytomegalovirus (HCMV) and being useful for the production of reconstituted HCMV virus retaining phenotypic characteristics of a clinical virus isolate including the ability to grow on endothelial cells and to induce microfusion is characterized in that it is obtainable by inserting DNA from a clinical isolate of HCMV virus into a bacterial cloning vehicle. Such vector can be used e.g., for production of reconstituted HCMV virus retaining the phenotypic characteristics of a parental clinical isolate and for studying genes and functions of genes of HCMV virus. A further aspect are mutant viruses and inter alia their use for studying aspects of infectivity of HCMV virus.

1 . Recombinant vector containing infectious genome of human cytomegalovirus (HCMV) and being useful for the production of reconstituted HCMV virus retaining phenotypic characteristics of a clinical virus isolate including the ability to grow on endothelial cells, to induce microfusion and to spread viral material from HUVEC to PMNL, characterized in that it is obtainable by inserting DNA from a clinical isolate of HCMV virus into a bacterial cloning vehicle. 
     
     
         2 . Recombinant vector according to  claim 1  which contains the complete infectious DNA of HCMV. 
     
     
         3 . Recombinant vector according to  claim 1 , characterized in that the bacterial cloning vehicle contains DNA sequences which are homologous to the HCMV DNA and insertion is effected by homologous recombination. 
     
     
         4 . Recombinant vector according to  claim 1 , wherein the bacterial cloning vehicle is a BAC system vector. 
     
     
         5 . Recombinant vector according to  claim 1  and designated FIX-Bac-7. 
     
     
         6 . Bacterial cell line containing a recombinant vector according to  claim 1 . 
     
     
         7 . Bacterial cell line according to  claim 6  which is an  E. coli  cell line. 
     
     
         8 . Bacterial cell line according to  claim 6 , designated FIX-Bac-7- E. coli  DH 10B and deposited as DSM 13958 containing at least one copy of FIX-Bac-7. 
     
     
         9 . A bacterial cell line according to  claim 6  for the production of reconstituted HCMV Virus retaining the phenotypic characteristics of a parental clinical isolate including the ability to grow on endothelial cells, to induce microfusion and to spread viral material from HUVEC to PMNL.