Confirmation of Transformation and DNA as the Inducing Material
The experiments in this report are a recreation the famous Griffith and Avery experiments which discovered the transformation process that bacteria can undertake and that DNA is the genetic material, respectively (Griffth, 1928. Avery 1944). Bacteria can incorporate foreign DNA found in their medium into their genomes in a process known as transformation. This process can be accelerated in CaCl solution at colder temperatures. The first part of this report deals with confirming that transformation does occur following a standard procedure, and the second of the report uses a similar procedure to confirm that DNA is the particle that induces transformation.
In the following procedures Escherichia coli was used:
(Part 1) For the first set of experiments a solution of half live ampicillin sensitive strain and half of a heat killed ampicillin resistant strain was mixed and incubated at 37°C for forty five minutes. The resulting fluid was spread onto a prewarmed LB and a prewarmed LB Amp100 medium, respectively. In addition six controls were set up which shared a LB and a LB Amp100 medium. These controls were given their own area on the media and streak plated. The controls consisted of the live ampicillin sensitive strain, the dead ampicillin resistant strain and the live ampicillin resistant strain plated onto each medium. Aseptic technique was used in treating all solutions, media and transfers between.
(Part 2) In the second set of experiments 1.0µL of kanamycin sensitive bacteria was added to two test tubes, both subjected to the same treatment. The bacteria in the...
... middle of paper ...
...colonies for live ampicillin sensitive cells grown on LB Amp100 media with only the buffer.
The colonies found on media treated with both the live ampicillin sensitive and heat killed resistant bacteria generated colonies with one exception: The treatment which used DNAse. This treatment showed the same result as the ones without heat killed bacteria: no growth was observed indicating that transformation could not occur without DNA. In conclusion this confirms that DNA is the hereditary medium which can transform bacteria, as discovered by Avery in 1944.
Avery, O. T., MacLeod, C. M. & McCarty, M. (1944). Studies on the Chemical Nature of the Substance Inducing Transformation of Pneumococcal Types. The Journal of Experimental Medicine, 79, 137-158.
Griffith F. (1928). The Significance of Pneumococcal Types. Epidermiology & Infection, 27(2), 113-159.
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