Genes in the HindIII J fragment of the murine cytomegalovirus genome are dispensable for growth in cultured cells: insertion mutagenesis with a lacZ/gpt cassette.
The organization and function of the genes encoded within the HindIII J region of the murine cytomegalovirus genome were analyzed by transcript mapping and cDNA isolation, nucleotide sequence analysis and identification of open reading frames (ORFs), and construction of recombinant viruses carrying insertions disrupting five of the seven ORFs. This region was found to encode five beta transcripts and one gamma transcript in addition to two beta transcripts previously mapped to the sgg1 locus. Seven open reading frames were identified, and one was recognized as a homolog of a human cytomegalovirus US22 gene family. The five largest ORFs contained within the HindIII J fragment (sgg1, HJ4, HJ5, HJ6, and HJ7) were each disrupted by the insertion of a lacZ/gpt genetic marker cassette. The growth kinetics of all recombinant viruses were investigated and found to be the same as wild-type parental virus, indicating that these five ORFs were dispensable for growth in cell culture.
Vieira
J
J
Department of Microbiology and Immunology, Stanford University School of Medicine, California 94305-5402.
Farrell
H E
HE
Rawlinson
W D
WD
Mocarski
E S
ES
D016475:3T3 Cells; D000818:Animals; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics* / Q000502:physiology; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D005814:Genes, Viral; D007700:Kinetics; D051379:Mice; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014779:Virus Replication / Q000235:genetics*
D015247:Deoxyribonuclease HindIII