The effects of transcriptional regulatory sequences introduced into a retroviral genome.

Chimeric plasmids in which the herpes thymidine kinase (tk) gene replaced portions of the Rous sarcoma viral genome were used to assess the relationship between viral transcription and that of an exogenous gene located within the viral genome. The entire tk gene and portions of the gene were positioned in both orientations within the viral gag and pol genomic region (which serves as intron for viral env mRNA). Microinjection assays then determined the amount of viral genomic transcription by quantitation of the amount of viral env mRNA produced. Separate injections also assayed for the presence of tk mRNA. Both mRNAs were produced unless the 3' region of the tk gene was present within the viral genome and in the same transcriptional sense. In this case viral env mRNA production was nearly abolished.

[1]  S. Pestka,et al.  Use of Rous sarcoma viral genome to express human fibroblast interferon. , 1986, Methods in enzymology.

[2]  I. Verma,et al.  Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene , 1985, Molecular and cellular biology.

[3]  M. Emerman,et al.  Genes with promoters in retrovirus vectors can be independently suppressed by an epigenetic mechanism , 1984, Cell.

[4]  H. Weintraub,et al.  Inhibition of thymidine kinase gene expression by anti-sense RNA: A molecular approach to genetic analysis , 1984, Cell.

[5]  M. Emerman,et al.  High-frequency deletion in recovered retrovirus vectors containing exogenous DNA with promoters , 1984, Journal of virology.

[6]  B. Cullen,et al.  Transcriptional interference in avian retroviruses—implications for the promoter insertion model of leukaemogenesis , 1984, Nature.

[7]  D. Stacey,et al.  Selective ligation of DNA molecules following microinjection. , 1983, The Journal of biological chemistry.

[8]  B. Cullen,et al.  Effect of intron size on splicing efficiency in retroviral transcripts. , 1982, Nucleic acids research.

[9]  D. Stacey,et al.  Analysis by microinjection of the biological effects of site-directed mutagenesis in cloned avian leukosis viral DNAs , 1982, Journal of virology.

[10]  H. Varmus,et al.  The leader sequence of the subgenomic mRNA's of Rous sarcoma virus is approximately 390 nucleotides , 1982, Journal of virology.

[11]  H. Temin,et al.  Formation of infectious progeny virus after insertion of herpes simplex thymidine kinase gene into DNA of an avian retrovirus , 1981, Cell.

[12]  E. Scolnick,et al.  Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1 , 1981, Journal of virology.

[13]  A. Skalka,et al.  Biological activity of cloned retroviral DNA in microinjected cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[14]  D. Stacey.  Microinjection of mRNA and other macromolecules into living cells. , 1981, Methods in enzymology.

[15]  Mario R. Capecchi,et al.  High efficiency transformation by direct microinjection of DNA into cultured mammalian cells , 1980, Cell.

[16]  H. Hanafusa,et al.  Microinjection analysis of envelope-glycoprotein messenger activities of avian leukosis viral RNAs. , 1977, Proceedings of the National Academy of Sciences of the United States of America.