2016 Research in the Capitol

Title

Optimizing Sucrose Gradients for Small Bacteriophage Purification

Presentation Type

Poster Presentation

Keywords

Bacteriophages--Morphology;

Abstract

Bacteriophages are viruses of bacteria and are abundant in nature. These viruses have differing morphologies; some have a head-tail morphology, while others have a tail-less morphology. Tail-less phages are under represented in the scientific literature and have properties of interest for certain applications. Current laboratory methods for isolating bacteriophages, however, have a strong tendency to select for larger phages with head-tail structure. The goal of this project was therefore to refine an existing method for bulk isolation of small bacteriophages from soil via sucrose density gradients. After centrifugation, these gradients were fractionated and spotted on bacterial lawns to assess infectivity. Fractions corresponding to peaks of infectivity were subject to SDS-PAGE to determine the presence of small phages. Sucrose gradients of 10-20% resolved phages better than 10-40% gradients. Finally, greater resolution of small phages was achieved with shorter periods of centrifugation.

Start Date

29-3-2016 11:30 AM

End Date

29-3-2016 1:30 PM

Event Host

University Honors Programs, Iowa Regent Universities

Faculty Advisor

Michael Walter

Department

Department of Biology

Comments

Location: Iowa State House, Rotunda, Des Moines, Iowa

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Mar 29th, 11:30 AM Mar 29th, 1:30 PM

Optimizing Sucrose Gradients for Small Bacteriophage Purification

Bacteriophages are viruses of bacteria and are abundant in nature. These viruses have differing morphologies; some have a head-tail morphology, while others have a tail-less morphology. Tail-less phages are under represented in the scientific literature and have properties of interest for certain applications. Current laboratory methods for isolating bacteriophages, however, have a strong tendency to select for larger phages with head-tail structure. The goal of this project was therefore to refine an existing method for bulk isolation of small bacteriophages from soil via sucrose density gradients. After centrifugation, these gradients were fractionated and spotted on bacterial lawns to assess infectivity. Fractions corresponding to peaks of infectivity were subject to SDS-PAGE to determine the presence of small phages. Sucrose gradients of 10-20% resolved phages better than 10-40% gradients. Finally, greater resolution of small phages was achieved with shorter periods of centrifugation.