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Team:Imperial College London/M3/DamMethylation
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=[[Image:II09_Thumb_m3.png|40px]]<font size='5'><b>Module 3: Genome Deletion Overview</b></font>= | =[[Image:II09_Thumb_m3.png|40px]]<font size='5'><b>Module 3: Genome Deletion Overview</b></font>= | ||
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Revision as of 13:58, 8 October 2009
Module 3: Genome Deletion Overview
Dam methylation
To protect against DNA destruction due to basal levels of restriction enzyme production, we have made use of the native E. coli Dam methylase protection system.
Dam methylases recognise the sequence GATC and methylate the Adenine base. This prevents the restriction enzymes from recognising the sequence and cleaving it. Therefore, only high levels of restriction enzyme (ie. after thermal triggering) will cleave the DNA.
There is an asymmetry between restriction enzymes and methylases. Restriction enzymes can cause just one cleavage that kills the cell. However, to protct the cell, methylases need to methylate all the recognition sites.
The methylase protection system has been used with its native promoter to prevent basal levels of restriction enzyme expression from killing the cell. The use of a native promoter shows the highly sensitive balance that exist between restriction enzymes and methylases.
