Methodology in Basic Genetics

By Walter J. Burdette | Go to book overview

Robert S. Edgar, Ph.D.


SOME TECHNICAL CONSIDERATIONS CONCERNING EXPERIMENTS on PHAGE RECOMBINATION

This discussion, meant primarily for the beginning worker in phage genetics, presupposes some knowledge of phage techniques and some elementary knowledge of genetics. It is meant to be more of a travel guide than a cookbook, helping the novice to avoid some of the more common errors and difficulties encountered in studies on phage recombination. For necessary information about basic techniques used in handling phage, the reader is referred to Adams' excellent manual.2

While many types of bacteriophage are known, only certain of those which attack Escherichia coli, the so-called coliphages T1,90, 91, 92T2,381, 538, 1026T4,212, 213 and lambda419, 450, 954, 1085 have been studied extensively from a genetic point of view, although a few studies on a smaller scale have been made on other phages such as a Salmonella phage, P22,533 and another coliphage, S13.1007 Since each phage is handled somewhat differently, by methods which have usually been developed empirically, T4 will be used as a general example along with methods used in the author's laboratory. Special features of other phages will be mentioned in passing. Hopefully the reader can gain a general idea of genetic methods used for bacteriophage and fill in details by further reading. In studying recombination in bacteriophage, certain procedures basic to genetic studies with any organism are followed, to make stable, genetically homogeneous stocks (populations). Different genotypes must be scored, and suitable mutants, either of spontaneous or induced origin, must be isolated. Finally, a standardized method of crossing different genotypes must be mastered, in order to do reproducible mapping. A phage cross consists of infecting a population of susceptible bacteria with phage of two or more genotypes, so that each bacterium has a mixed

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