Stable Insertion into the Human Cytomegalovirus Genome of a Foreign Gene.

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On the bases of previous finding that the HindIII O fragment of 7.8-kb in the genome of human cytomegalovirus strain Towne is nonessential for viral replication, we constructed a vector pKM that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using the vector, we constructed strain Towne-derived recombinant viruses in which a chimeric lacZ gene fused to the SV40 promoter and poly A signals was inserted in place of the O fragment. Two types of recombinants were obtained which carry the chimeric gene in opposite directions. Beta Gal was expressed in HEL cells infected with these recombinants throughout the infection cycle, and the rate of its synthesis in early stages of infection was comparable to that of synthesis of a 65 kDa viral glycoprotein, one of viral proteins expressed abundantly. The chimeric lacZ gene introduced was stable and no lacZ- revertant has been observed so far.