Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1981 Jan;37(1):343–351. doi: 10.1128/jvi.37.1.343-351.1981

Genetic analysis of gene 1.2 of bacteriophage T7: isolation of a mutant of Escherichia coli unable to support the growth of T7 gene 1.2 mutants.

H Saito, C C Richardson
PMCID: PMC171012  PMID: 7012382

Abstract

The product of gene 1.2 of bacteriophage T7 is not required for the growth of T7 in wild-type Escherichia coli since deletion mutants lacking the entire gene 1.2 grow normally (Studier et al., J. Mol. Biol. 135:917-937, 1979). By using a T7 strain lacking gene 1.2, we have isolated a mutant of E. coli that was unable to support the growth of both point and deletion mutants defective in gene 1.2. The mutation, optA1, was located at approximately 3.6 min on the E. coli linkage map in the interval between dapD and tonA; optA1 was 92% cotransducible with dapD. By using the optA1 mutant, we have isolated six gene 1.2 point mutants of T7, all of which mapped between positions 15 and 16 on the T7 genetic map. These mutations have also been characterized by DNA sequence analysis, E. coli optA1 cells infected with T7 gene 1.2 mutants were defective in T7 DNA replication; early RNA and protein synthesis proceeded normally. The defect in T7 DNA replication is manifested by a premature cessation of DNA synthesis and degradation of the newly synthesized DNA. The defect was not observed in E. coli opt+ cells infected with T7 gene 1.2 mutants or in E. coli optA1 cells infected with wild-type T7 phage.

Full text

PDF
343

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benbasat J. A., Burck K. B., Miller R. C., Jr Superinfection exclusion and lack of conservative transfer of bacteriophage T7 DNA. Virology. 1978 Jun 1;87(1):164–171. doi: 10.1016/0042-6822(78)90168-x. [DOI] [PubMed] [Google Scholar]
  3. Chase J. W., Richardson C. C. Escherichia coli mutants deficient in exonuclease VII. J Bacteriol. 1977 Feb;129(2):934–947. doi: 10.1128/jb.129.2.934-947.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Lucia P., Cairns J. Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1164–1166. doi: 10.1038/2241164a0. [DOI] [PubMed] [Google Scholar]
  5. DeWyngaert M. A., Hinkle D. C. Bacterial mutants affecting phage T7 DNA replication produce RNA polymerase resistant to inhibition by the T7 gene 2 protein. J Biol Chem. 1979 Nov 25;254(22):11247–11253. [PubMed] [Google Scholar]
  6. Gellert M., Bullock M. L. DNA ligase mutants of Escherichia coli. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1580–1587. doi: 10.1073/pnas.67.3.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  8. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Masamune Y., Frenkel G. D., Richardson C. C. A mutant of bacteriophage T7 deficient in polynucleotide ligase. J Biol Chem. 1971 Nov 25;246(22):6874–6879. [PubMed] [Google Scholar]
  10. Oakley J. L., Coleman J. E. Structure of a promoter for T7 RNA polymerase. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4266–4270. doi: 10.1073/pnas.74.10.4266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Panayotatos N., Wells R. D. Identification, cloning and characterization of three late promoters at 14.6, 14.8 and 15.9% of T7 DNA. J Mol Biol. 1979 Nov 25;135(1):91–109. doi: 10.1016/0022-2836(79)90342-5. [DOI] [PubMed] [Google Scholar]
  12. Rahmsdorf H. J., Pai S. H., Ponta H., Herrlich P., Roskoski R., Jr, Schweiger M., Studier F. W. Protein kinase induction in Escherichia coli by bacteriophage T7. Proc Natl Acad Sci U S A. 1974 Feb;71(2):586–589. doi: 10.1073/pnas.71.2.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Richardson C. C., Romano L. J., Kolodner R., LeClerc J. E., Tamanoi F., Engler M. J., Dean F. B., Richardson D. S. Replication of bacteriophage T7 DNA by purified proteins. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):427–440. doi: 10.1101/sqb.1979.043.01.049. [DOI] [PubMed] [Google Scholar]
  14. Richardson C. C. The 5'-terminal nucleotides of T7 bacteriophage deoxyribonucleic acid. J Mol Biol. 1966 Jan;15(1):49–61. doi: 10.1016/s0022-2836(66)80208-5. [DOI] [PubMed] [Google Scholar]
  15. Saito H., Tabor S., Tamanoi F., Richardson C. C. Nucleotide sequence of the primary origin of bacteriophage T7 DNA replication: relationship to adjacent genes and regulatory elements. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3917–3921. doi: 10.1073/pnas.77.7.3917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Saito H., Uchida H. Initiation of the DNA replication of bacteriophage lambda in Escherichia coli K12. J Mol Biol. 1977 Jun 15;113(1):1–25. doi: 10.1016/0022-2836(77)90038-9. [DOI] [PubMed] [Google Scholar]
  17. Shizuya H., Livingston D. M., Richardson C. C. Isolation of a specialized transducing bacteriophage lambda carrying the polC locus of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2614–2617. doi: 10.1073/pnas.71.7.2614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Simon M. N., Studier F. W. Physical mapping of the early region of bacteriophage T7 DNA. J Mol Biol. 1973 Sep 15;79(2):249–265. doi: 10.1016/0022-2836(73)90004-1. [DOI] [PubMed] [Google Scholar]
  19. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  20. Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
  21. Studier F. W. Gene 0.3 of bacteriophage T7 acts to overcome the DNA restriction system of the host. J Mol Biol. 1975 May 15;94(2):283–295. doi: 10.1016/0022-2836(75)90083-2. [DOI] [PubMed] [Google Scholar]
  22. Studier F. W. Genetic analysis of non-essential bacteriophage T7 genes. J Mol Biol. 1973 Sep 15;79(2):227–236. doi: 10.1016/0022-2836(73)90002-8. [DOI] [PubMed] [Google Scholar]
  23. Studier F. W., Rosenberg A. H., Simon M. N., Dunn J. J. Genetic and physical mapping in the early region of bacteriophage T7 DNA. J Mol Biol. 1979 Dec 25;135(4):917–937. doi: 10.1016/0022-2836(79)90520-5. [DOI] [PubMed] [Google Scholar]
  24. Studier F. W. The genetics and physiology of bacteriophage T7. Virology. 1969 Nov;39(3):562–574. doi: 10.1016/0042-6822(69)90104-4. [DOI] [PubMed] [Google Scholar]
  25. Tamanoi F., Saito H., Richardson C. C. Physical mapping of primary and secondary origins of bacteriophage T7 DNA replication. Proc Natl Acad Sci U S A. 1980 May;77(5):2656–2660. doi: 10.1073/pnas.77.5.2656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wu T. T. A model for three-point analysis of random general transduction. Genetics. 1966 Aug;54(2):405–410. doi: 10.1093/genetics/54.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES