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PMID:3011596
| Citation |
Morris, CE, Klement, JF and McAllister, WT (1986) Cloning and expression of the bacteriophage T3 RNA polymerase gene. Gene 41:193-200 |
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| Abstract |
The gene that encodes the RNA polymerase of bacteriophage T3 (gene 1) has been cloned into a pBR322 derivative under the control of an inducible lacUV5 promoter. Large quantities of the protein are synthesized after induction of cells that carry this plasmid. RNA polymerase purified from these overproducing cells selectively initiates transcription from T3 promoter sequences as demonstrated by transcription of a dual promoter plasmid that carries both T3 and T7 promoters. Cells that carry the T3 RNA polymerase gene can complement amber mutants of T3 that are defective in gene 1 but not gene 1 amber mutants of T7, and vice versa; this experiment demonstrates the specificity of these enzymes in vivo. |
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| Keywords |
Base Sequence; Cloning, Molecular; DNA Restriction Enzymes; DNA-Directed RNA Polymerases/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Genes; Genes, Viral; Plasmids; Promoter Regions, Genetic; T-Phages/enzymology; T-Phages/genetics; Transcription, Genetic |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0003899: DNA-directed RNA polymerase activity |
ECO:0000314: |
F |
Fig 5. The T3 gene 1.0 was over-expressed on a plasmid, and the protein product was collected. T3 mutants with no functional gene 1.0 were incapable of initiating transcription, but regained function when in the presence of collected gp1.0 protein. T7 phages without a functional copy of gene 1.0 were not affected by phage T3 gp1.0. This suggests that the T3 gene 1.0 is a T3-specific RNA polymerase. |
complete | ||||
Notes
See also
References
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