Bacteria

The attraction of bacteria such as E. coli or B. subtil is as experimental systems is that they are relatively simple cells and can be grown and manipulated with comparative ease. Bacteria are single-celled organisms in which all of the machinery for DNA, RNA, and protein synthesis is contained in the same cellular compartment (bacteria have nn nucleus}.

Bacteria usually have a single chromosome—typically much smaller than the genome of higher organisms, Also, bacteria have a short generation time (the cell cycle can be as short as 20 minutes) and a genetically homogenous population of cells (a clone) can easily be generated from a single cell. Finally, bacteria are convenient to study genetically because, on the one hand, they are haploid (which means that the phenotypes of mutations, even recessive mutations, manifest readily), and, on the other hand, because genetic material can be conveniently exchanged between bacteria,

Molecular biology owes its origin to experiments with bacterial and phage model systems. Up until the famous fluctuation analysis experiments of Salvadore Luria and Max Delruck in 1943, the study of bacteria (bacteriology) had remained largely outside the realm of traditional genetics. Taking a statistical approach, Luria and Delruck demonstrated that bacteria can undergo a change in which they become resistant to infection by a particular phage. Critically, they showed that this change arises spontaneously, rather than as a response (adaptation) to the phage. Thus, like other organisms, bacteria can inherit traits (for example, sensitivity or resistance to a phage), and occasionally this inheritance can undergo a spontaneous change (mutation) to an alternative inheritable state. The experiments of Luria and Delruck showed that, like other organisms, bacteria exhibit genetically determined characteristics. But because of their simplicity, bacteria would be ideal experimental systems in which to elucidate the nature of the genetic material and the trait-de-termining factors (genes) of Cregor Mendel.

Assays of Bacterial Growth

Bacteria can be grown in liquid or on solid (agar) medium. Bacterial cells are large enough (about 2 p.rn in length) to scatter light, allowing the growth of a bacterial culture to be monitored conveniently in liquid culture by the increase in optical density. Actively growing bacteria that are dividing with a constant generation time increase in numbers exponentially. They are said to be in the exponential phase of growth. As the population increases to high numbers of cells, the growth rate slows and bacteria enter the stationary phase (Figure 21-5).

The number of bacteria can be determined by diluting the culture and plating the cells on solid (agar) medium in a petri dish. Single cells grow into macroscopic colonies consisting of millions of cells within a relatively brief period of time. Knowing how many colonies are on the plate and how much the culture was diluted makes it possible to calculate the concentration or cells in the original culture.

Cj JD

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