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Identification and mapping of five new genes in bacteriophage T7
Authors:FWilliam Studier
Affiliation:Biology Department, Brookhaven National Laboratory Upton, N.Y. 11973, U.S.A.
Abstract:Mutations that affect the single-stranded DNA-binding protein of bacteriophage T7 (gene 2.5) and four T7 proteins of unknown function (the gene 4.3, 4.5, 4.7 and 5.5 proteins) are described and mapped by three-factor crosses. An extensive search for mutants defective in the DNA-binding protein (Mr = 25,562) produced several strains in which this protein has an altered electrophoretic mobility but no strains that appear to lack it completely. The gene 2.5 mutation that was mapped produces a slightly short DNA-binding protein that appears functional by tests in vitro. It seems likely that a functional DNA-binding protein is needed for T7 growth but that conditional-lethal amber mutations in it are rare; the nucleotide sequence known to code for the gene 2.5 protein contains only 1 to 3 sites that would be expected to be readily mutable to conditional-lethal amber codons by N-methyl-N?nitro-N-nitrosoguanidine. The gene 4.3, 4.5 and 4.7 proteins (Mr ~ 8000 to 15,000) are eliminated by a deletion mutant that removes most of the DNA between genes 4 and 5. The gene 5.5 protein (Mr ~ 11,700) is made in relatively large amounts and is affected by two different mutations that were mapped between genes 5 and 6. One of these mutations appears to be an amber mutation that eliminates the protein entirely; the other decreases the electrophoretic mobility of the protein (an apparent increase in size). A larger protein (Mr ~ 18,000), found in small amounts and difficult to observe, is also affected by these two mutations; the relationship of this minor protein to the major gene 5.5 protein is not yet known. The genes 2 and 18 proteins have also been identified in patterns of protein synthesis during infection. The proteins specified by at least 34 different T7 genes have now been identified.
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