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Cellular transcripts encoded at a locus which permits retrovirus expression in mouse embryonic cells.
R Petersen1, S Sobel, C T Wang
1Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, Portland 97201.
Gene
|May 30, 1991
Summary
Recombinant retroviruses activated in mouse F9 cells reveal the F2 locus. This conserved mammalian locus expresses a 450-nucleotide transcript, potentially encoding a constitutive function or a nonfunctional element.
Area of Science:
- Molecular Biology
- Genomics
- Mammalian Genetics
Background:
- The F2 locus in mouse embryonal carcinoma cells was investigated for its transcriptional activity and evolutionary conservation.
- Recombinant retroviruses were utilized to identify and characterize elements within the F2 locus.
Purpose of the Study:
- To elucidate the structure and expression of the F2 locus in mammalian cells.
- To determine the potential function of the F2 locus-encoded transcript.
Main Methods:
- Retroviral insertion mutagenesis in F9 cells.
- Transient transfection assays with chloramphenicol acetyltransferase reporter.
- Isolation and analysis of F2 cDNA clones.
- Sequence homology analysis across mammalian species.
Main Results:
- Proviral integration activated the F2 promoter, driving expression of a 450-nucleotide transcript in various cell types and tissues.
- F2 homologous sequences are conserved across mammalian genomes, with the 450-nt transcript expressed in rat and human cells.
- F2 cDNA clones revealed repeats and identified integration sites within introns, but lacked long open reading frames or significant homology to known sequences.
Conclusions:
- The F2 locus encodes a conserved 450-nucleotide transcript expressed in mammals.
- The F2 locus may represent a constitutively required function or a conserved, expressed, nonfunctional element.