In Vitro Genetic Recombination of Bacteriophage λ
- 1 June 1974
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 71 (6), 2496-2499
- https://doi.org/10.1073/pnas.71.6.2496
Abstract
DNA of bacteriophage lambda recombines in a cell-free extract prepared from an induced Escherichia coli lysogen of bacteriophage lambda. The assay for recombination in vitro takes advantage of the ability of such an extract to package lambda DNA and to assemble complete phage particles. For example, when lambda DNA that has been extracted from phage with the immunity of 434 is added to an extract, infectious lambda imm 434 particles are produced. The precursor DNA molecule in this packaging reaction is a multichromosomal polymer; circular monomers, for example, are not packaged.Nevertheless, when 434 circular DNA monomers are added to an extract, some phage that contain the imm 434 marker are produced. In this case, the circular DNA had recombined with lambda DNA in the extract and thereby had become part of a polymeric structure, which by the normal packaging process could give rise to infectious particles with the imm 434 marker. Genetic recombination is demonstrated when imm 434 circular monomer DNA carries amber mutations in genes A and B; then most of the 434 plaque formers produced in vitro are A(+)B(+), the genotype of the endogenous lambda DNA. Genetic crossing-over occurs through a region that contains the prophage attachment site, suggesting that recombination is carried out by the lambda Int functions. The 434 recombinant plaque formers are particles physically identical to wild-type 434 particles, as judged by their buoyant density in a CsCl equilibrium gradient.Keywords
This publication has 17 references indexed in Scilit:
- Deoxyribonucleic acid ligase. A steady state kinetic analysis of the reaction catalyzed by the enzyme from Escherichia coli.1973
- Deletion mutants of bacteriophage lambda: II. Genetic properties of att-defective mutantsJournal of Molecular Biology, 1971
- Properties of recombination-deficient mutants of bacteriophage lambdaJournal of Molecular Biology, 1970
- The role of exonuclease and β protein of bacteriophage λ in genetic recombination: I. Effects of red mutants on protein structureJournal of Molecular Biology, 1970
- Biochemical and Genetic Studies of Recombination Proficiency in Escherichia coli , I. Enzymatic Activity Associated with recB + and recC + GenesProceedings of the National Academy of Sciences, 1970
- Relations physiologiques entre les phages tempérés λ et Φ80Molecular Genetics and Genomics, 1970
- Degree of superhelicity of covalently closed cyclic DNA's from Escherichia coliJournal of Molecular Biology, 1969
- Vegetative λ DNAJournal of Molecular Biology, 1968
- Enzymatic DNA Degradation in E. coli: Its Relationship to Synthetic Processes at the Chromosome LevelCold Spring Harbor Symposia on Quantitative Biology, 1968
- Thermodynamic and kinetic studies on the interconversion between the linear and circular forms of phage lambda DNAJournal of Molecular Biology, 1966