Simple method for detecting inhibitors of retroviral replication

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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435 6, 4352402, 4352542, 4353201, C12Q 170, C12N 119, C12N 1581

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057143137

ABSTRACT:
The present invention relates, in general, to a DNA segment. In particular, the present invention relates to a DNA segment comprising a selectable marker gene, a DNA segment comprising a selectable marker gene inserted into a retrotransposon, cells containing these DNA segments, and methods of using these DNA segments and cells. The present invention further relates to a vector comprising a selectable marker gene inserted into a retrotransposon, wherein the retrotransposon comprises a retroviral reverse transcriptase/RNase H gene domain and wherein the selectable marker gene contains an intron inserted into a coding sequence of the marker gene and the intron is in antisense orientation relative to transcription of the marker gene and in sense orientation relative to transcription of the retrotransposon.

REFERENCES:
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Boeke, J.D. et al., "A General Method for the Chromosomal Amplification of Genes in Yeast" Science 239:280-282 (Jan. 1988).
Boeke, J.D. et al., "Ty Elements Transpose Through an RNA Intermediate" Cell 40:491-500 (Mar. 1985).
Heidmann, T. et al., "An Indicator Gene to Demonstrate Intracellular Transposition of Defective Retroviruses," Proc. Natl. Acad. Sci. USA 85:2219-2223 (Apr. 1988).
Kim and Loeb, "Human Immunodeficiency Virus Reverse Transcriptase Substitutes for DNA Polymerase I in Escherichia coli," Proc. Natl. Acad. Sci. USA 92:684-688, Jan. 1995.
Curcio and Garfinkel, "Single-Step Selection for TY1 Element Retrotransposition," Proc. Natl. Acad. Sci. USA 88:936-940 (Feb. 1991).
Dombroski et al., "An in Vivo Assay for the Reverse Transcriptase of Human Retrotransposon L1 in Saccharomyces cerevisiae," Molecular and Cellular Biology 14(7):4485-4492 (Jul. 1994) .
Sharon et al., "Efficient Homologous Recombination of Ty1 Element cDNA when Intergration is blocked," Molecular and Cellular Biology 14(10):6540-6551 (Oct. 1994).
Derr and Strathern, "A Role for Reverse Transcripts in Gene Conversion," Nature 361:170-173 (Jan. 14, 1993).
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