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W J Brammar

Publications and source records attributed to W J Brammar.

86 records · Page 5Linked to original sources
PubMed1976

The construction in vitro of transducing derivatives of phage lambda.

Methods are described for the construction of plaque-forming, transducing derivatives of phage lambda, using appropirate receptor genomes and fragments of DNA generated by the restriction enzymes endo R.EcoRI and endo R.HindIII. The general properties of the transducing derivatives are described and discussed. Plaque-forming phages carrying the E. coli trp, his, cysB, thyA, supD, supE, sup F, hsd, tna and lig genes have been isolated.

Chromosomes, Bacterial↗
PubMed1975

Punctuation of transcription in vitro of the tryptophan operon of Escherichia coli. A novel type of control of transcription.

RNA transcribed in vitro from DNA of a tryptophan (trp) transducing strain of bacteriophage phi80 which contains the trp regulatory elements consists of a polycistronic messenger transcribed from the structural genes, and possibly the regulatory region, and a separate RNA species (called trp regRNA) which is transcribed from the regulatory region. This conclusion is based on hybridization experiments with trp RNA synthesized in vitro and the separate DNA strands of trp transducing strains of lambda with and without the trp regulatory elements. The length of trp regRNA determined by filtration on Sephadex G-200 is 110-180 nucleotides. From the amount and the length of trp regRNA we have calculated that 8-20 copies of trp regRNA are synthesized per copy of polycistronic trp mRNA. We conclude that during transcription of the trp operon RNA polymerase frequently is rejected at a specific site ahead of the first structural gene, trpE. The termination factor Rho is not involved in this process. A different protein fraction, which specifically stimulates the synthesis of trp enzymes in an in vitro protein-synthesizing system (Pouwels and Van Rotterdam, 1975), was found to antagonize the abortive synthesis of trp mRNA. A model is proposed for the control of transcription of the trp genes, which operates through a mechanism of punctuation of RNA synthesis at a specific site on the DNA template and anti-termination of RNA synthesis by means of a positive control factor.

Coliphages↗
PubMed1968

Spontaneous and ICR191-A-induced frameshift mutations in the A gene of Escherichia coli tryptophan synthetase.

Frameshift mutant trpA21 was isolated after ultraviolet treatment and frameshift mutant trpA540 after ICR191-A (an acridine derivative) treatment of wild-type Escherichia coli K-12. The A proteins of spontaneous and ICR191-A-induced partial revertants of these mutants contained altered amino acid sequences one residue shorter than the comparable sequence in the A protein of wild-type bacteria. The data support the conclusion that ICR191-A causes frameshift mutations. The findings further indicate that both base additions and deletions are elicited by ICR191-A treatment and that mutagenesis by this compound sometimes affects more than one base pair. ICR191-A also weakly reverts some missense mutants. Analyses of the relevant peptides of the purified A protein show single amino acid replacements compatible with single base-pair changes. In addition, we found that some spontaneously revertible ICR191-A- and ultraviolet light-induced frameshift mutants are not further stimulated to revert by exposure to ultraviolet light.

Acridines↗