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. 1987 Oct;169(10):4630–4636. doi: 10.1128/jb.169.10.4630-4636.1987

Mutations in an upstream regulatory sequence that increase expression of the bacteriophage T4 lysozyme gene.

J A Knight 1, L W Hardy 1, D Rennell 1, D Herrick 1, A R Poteete 1
PMCID: PMC213832  PMID: 3654580

Abstract

A P22 hybrid phage bearing the bacteriophage T4 lysozyme gene (e), as well as T4 sequences upstream from the lysozyme gene, was constructed. Amber mutations were introduced into gene e in the hybrid phage, and the resulting mutant phages were tested for the ability to form plaques on amber suppressor strains. Revertant phages that were able to form plaques on amber suppressors that did not suppress the parent amber mutant phages were isolated following UV mutagenesis. Secondary site pseudorevertants were identified among the revertants by a genetic test. Four of the suppressing secondary site mutations were mapped and sequenced. They were found to consist of small sequence alterations immediately upstream from gene e, all of which would tend to destabilize potential base-pairing interactions in the transcript. The mutations were shown to increase lysozyme expression when introduced into an otherwise wild-type hybrid phage, but were found to have little effect on transcription of the lysozyme gene.

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Selected References

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  1. Berget P. B., Poteete A. R., Sauer R. T. Control of phage P22 tail protein expression by transcription termination. J Mol Biol. 1983 Mar 15;164(4):561–572. doi: 10.1016/0022-2836(83)90050-5. [DOI] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Graña D., Youderian P., Susskind M. M. Mutations that improve the ant promoter of Salmonella phage P22. Genetics. 1985 May;110(1):1–16. doi: 10.1093/genetics/110.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grütter M. G., Matthews B. W. Amino acid substitutions far from the active site of bacteriophage T4 lysozyme reduce catalytic activity and suggest that the C-terminal lobe of the enzyme participates in substrate binding. J Mol Biol. 1982 Jan 25;154(3):525–535. doi: 10.1016/s0022-2836(82)80011-9. [DOI] [PubMed] [Google Scholar]
  6. Inoue T., Cech T. R. Secondary structure of the circular form of the Tetrahymena rRNA intervening sequence: a technique for RNA structure analysis using chemical probes and reverse transcriptase. Proc Natl Acad Sci U S A. 1985 Feb;82(3):648–652. doi: 10.1073/pnas.82.3.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  8. McPheeters D. S., Christensen A., Young E. T., Stormo G., Gold L. Translational regulation of expression of the bacteriophage T4 lysozyme gene. Nucleic Acids Res. 1986 Jul 25;14(14):5813–5826. doi: 10.1093/nar/14.14.5813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Owen J. E., Schultz D. W., Taylor A., Smith G. R. Nucleotide sequence of the lysozyme gene of bacteriophage T4. Analysis of mutations involving repeated sequences. J Mol Biol. 1983 Apr 5;165(2):229–248. doi: 10.1016/s0022-2836(83)80255-1. [DOI] [PubMed] [Google Scholar]
  10. Perry L. J., Heyneker H. L., Wetzel R. Non-toxic expression in Escherichia coli of a plasmid-encoded gene for phage T4 lysozyme. Gene. 1985;38(1-3):259–264. doi: 10.1016/0378-1119(85)90226-4. [DOI] [PubMed] [Google Scholar]
  11. Rao G. R., Burma D. P. Purification and properties of phage P22-induced lysozyme. J Biol Chem. 1971 Nov;246(21):6474–6479. [PubMed] [Google Scholar]
  12. Remington S. J., Anderson W. F., Owen J., Ten Eyck L. F., Grainger C. T., Matthews B. W. Structure of the lysozyme from bacteriophage T4: an electron density map at 2.4 A resolution. J Mol Biol. 1978 Jan 5;118(1):81–98. doi: 10.1016/0022-2836(78)90245-0. [DOI] [PubMed] [Google Scholar]
  13. Rennell D., Poteete A. R. Phage P22 lysis genes: nucleotide sequences and functional relationships with T4 and lambda genes. Virology. 1985 May;143(1):280–289. doi: 10.1016/0042-6822(85)90115-1. [DOI] [PubMed] [Google Scholar]
  14. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  15. Weaver L. H., Rennell D., Poteete A. R., Mathews B. W. Structure of phage P22 gene 19 lysozyme inferred from its homology with phage T4 lysozyme. Implications for lysozyme evolution. J Mol Biol. 1985 Aug 20;184(4):739–741. doi: 10.1016/0022-2836(85)90318-3. [DOI] [PubMed] [Google Scholar]
  16. Weinstock G. M., Susskind M. M., Botstein D. Regional specificity of illegitimate recombination by the translocatable ampicillin-resistance element Tn1 in the genome of phage P22. Genetics. 1979 Jul;92(3):685–710. doi: 10.1093/genetics/92.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Winston F., Botstein D., Miller J. H. Characterization of amber and ochre suppressors in Salmonella typhimurium. J Bacteriol. 1979 Jan;137(1):433–439. doi: 10.1128/jb.137.1.433-439.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Youderian P., Bouvier S., Susskind M. M. Sequence determinants of promoter activity. Cell. 1982 Oct;30(3):843–853. doi: 10.1016/0092-8674(82)90289-6. [DOI] [PubMed] [Google Scholar]
  19. Youderian P., Susskind M. M. Identification of the products of bacteriophage P22 genes, including a new late gene. Virology. 1980 Nov;107(1):258–269. doi: 10.1016/0042-6822(80)90291-3. [DOI] [PubMed] [Google Scholar]
  20. Zagursky R. J., Berman M. L. Cloning vectors that yield high levels of single-stranded DNA for rapid DNA sequencing. Gene. 1984 Feb;27(2):183–191. doi: 10.1016/0378-1119(84)90139-2. [DOI] [PubMed] [Google Scholar]
  21. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]

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