Lidstrom:UV Mutagenesis: Difference between revisions
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== Basics of UV mutagenesis == | == Basics of UV mutagenesis == | ||
* In C. elegans, UV mutagenesis is better than EMS mutagenesis for large deletions and rearrangements. ([http://www.ncbi.nlm.nih.gov/pubmed/?term=8531732 Anderson 1995]) | * In C. elegans, UV mutagenesis is better than EMS mutagenesis for large deletions and rearrangements. ([http://www.ncbi.nlm.nih.gov/pubmed/?term=8531732 Anderson 1995]) | ||
** Note, however, the effect of mutagens is different on different hosts so this may not be true for your bacterium. | ** Note, however, the effect of mutagens is different on different hosts so this may not be true for your bacterium. | ||
== Reasons AM1 has been mutated with EMS, not UV == | |||
* The genome is GC rich. | |||
** Since UV makes TT dimers, you get less mutations and they are more frequently right before genes where the DNA tends to be AT rich. | |||
* It is pink, so it absorbs the UV radiation to some degree. | |||
** [[Users:Janet B. Matsen|JM]] doesn't think this should matter much. It should just mean that you need a larger dose. | |||
== Resources == | == Resources == | ||
Latest revision as of 16:14, 14 July 2014
Back to Protocols
Basics of UV mutagenesis
- In C. elegans, UV mutagenesis is better than EMS mutagenesis for large deletions and rearrangements. (Anderson 1995)
- Note, however, the effect of mutagens is different on different hosts so this may not be true for your bacterium.
Reasons AM1 has been mutated with EMS, not UV
- The genome is GC rich.
- Since UV makes TT dimers, you get less mutations and they are more frequently right before genes where the DNA tends to be AT rich.
- It is pink, so it absorbs the UV radiation to some degree.
- JM doesn't think this should matter much. It should just mean that you need a larger dose.
Resources
- UV Radiation and Spontaneous Mutagenesis: summary by Kendric C. Smith, Emeritus Professor, Radiation Oncology (Radiation Biology), Stanford University School of Medicine
- Discusses repair mechanisms a lot.
- UV radiation produces a preponderance of alterations in the pyrimidines, and the large majority of these products involve the linking of two adjacent pyrimidines
- "G:C -> A:T transitions predominate after UV irradiation.
- In E. coli the (6-4)-photoproduct may be more important for mutagenesis, while the pyrimidine dimer may be more important in mammalian cells. In human cells, mutations occur at the C of a TC, CT, or CC pyrimidine dimer, but not at TT dimers, and also occur at the C of TC and CC (6-4)-adducts (reviewed by Brash, 1988)."
- UV-radiation-induced mutations show 2-hit kinetics at high doses
- i.e., they are produced as a function of the square of the dose
- While UV irradiation does not produce DNA double-strand breaks directly, they can be formed as a consequence of the inefficient repair of overlapping excision gaps, and of overlapping daughter-strand gaps
- The formation of DNA-protein cross-links has been shown to be of significance in the killing of E. coli
- In general, UV irradiation produces mutations along a gene in a non-random manner, i.e., mutations are observed at certain base pairs more frequently than at others
