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Selected Topics from Classical Bacterial Genetics

Elisabeth A. Raleigh¹, Karen Elbing², Roger Brent³

¹New England Biolabs, Beverly, Massachusetts
²Clark & Elbing LLP, Boston, Massachusetts
³The Molecular Sciences Institute, Berkeley, California

Publication Name:

Current Protocols in Molecular Biology

Unit Number:

Unit 1.4

DOI:

10.1002/0471142727.mb0104s59

Online Posting Date:

August, 2002

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Abstract

Current cloning technology exploits many facts learned from classical bacterial genetics. This unit covers those that are critical to understanding the techniques described in this book. Topics include antibiotics, the LAC operon, the F factor, nonsense suppressors, genetic markers, genotype and phenotype, DNA restriction, modification and methylation and recombination.

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Unit Introduction
Antibiotics
The Lac Operon
The F Factor
Nonsense Suppressors
Genetic Markers
Genotype and Phenotype
DNA Restriction, Modification, and Methylation
Recombination and Its Effects on Cloned DNA Inserts
Effects of Recombination-Defective Strains on Vectors
Bibliography
Figures
Tables

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Figures

Figure 1.4.1

The first line shows the genes of the lac operon. lacZ, lacY, and lacA transcription is repressed by lac repressor. In the next line, inactive repressor is no longer bound to the operator, and the genes are being transcribed by RNA polymerase. In the third line, RNA polymerase transcribes the operon. Finally, RNA polymerase encounters a transcription terminator and, in a reaction requiring rho protein, RNA polymerase is detached from the DNA and the nascent transcript.

View Image
Figure 1.4.2

Alpha-complementation.

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Figure 1.4.3

Recombination between homologous direct repeats.

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Literature Cited

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Key References
	Beckwith, J. and Zipser, D. eds. 1970. The Lactose Operon. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y
An introduction to early work with the lactose operon.
	Miller, 1972. See above
Contains excellent introductions to experimental techniques for working with E. coli and -derived phages.
	Miller, J., ed. 1981. The Operon. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y
An updated and thorough introduction to regulatory mechanisms, starting with the lactose operon.
	Neidhardt, F.C., Ingraham, J.L., Low, K.B., Magasanik, B., Schaechter, M., and Umbarger, H.E., eds. 1996. Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology. American Society for Microbiology, Washington, D.C
Encyclopedic coverage of the biology of these useful bacteria.
	Ullman, A. and Perrin, D. 1970. Complementation in -galactosidase. In The Lactose Operon (J. Beckwith and D. Zipser, eds.) pp. 143-172. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y
Definitive review of alpha-complementation.