Submission Type
Event
Expected Graduation Date
2014
Location
Center for Natural Sciences, Illinois Wesleyan University
Start Date
4-12-2014 2:00 PM
End Date
4-12-2014 3:00 PM
Disciplines
Biology
Abstract
Endonucleases are enzymes which cleave DNA at specific nucleotide sequences. Homing Endonucleases (HEase) are a group of endonucleases that reside as open reading frames within self-splicing introns or inteins. After expression of the gene containing the HEase the HEase goes on to cleave a target site in a homolog of the hosting gene to induce homologous recombination turning the vacant homolog into a homing endonuclease gene (HEG). It is predicted that gene product (gp126) from the mycobacteriophage Gizmo encodes a HEase. To test whether the gene encodes a HEase the phage sequence of gp126 Gizmo was transferred into the his-tag containing plasmid (PET14b). The protein has been expressed and purified. To test the activity of the HEase a substrate is being created that will allow the predicted function to be directly tested in an enzyme assay.
Included in
Expression of Gene Product 126 from Phage Gizmo and Creation of a Substrate
Center for Natural Sciences, Illinois Wesleyan University
Endonucleases are enzymes which cleave DNA at specific nucleotide sequences. Homing Endonucleases (HEase) are a group of endonucleases that reside as open reading frames within self-splicing introns or inteins. After expression of the gene containing the HEase the HEase goes on to cleave a target site in a homolog of the hosting gene to induce homologous recombination turning the vacant homolog into a homing endonuclease gene (HEG). It is predicted that gene product (gp126) from the mycobacteriophage Gizmo encodes a HEase. To test whether the gene encodes a HEase the phage sequence of gp126 Gizmo was transferred into the his-tag containing plasmid (PET14b). The protein has been expressed and purified. To test the activity of the HEase a substrate is being created that will allow the predicted function to be directly tested in an enzyme assay.