Graduation Year

2014

Location

Center for Natural Sciences, Illinois Wesleyan University

Start Date

12-4-2014 2:00 PM

End Date

12-4-2014 3:00 PM

Description

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.

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Biology Commons

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Apr 12th, 2:00 PM Apr 12th, 3:00 PM

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.