Title

Visualizing the Pathways of MBP-EGFP Fusions with Fluorescence Microscopy

Poster Number

58

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Geoff Lin-Cereghino

Faculty Mentor Department

Biological Sciences

Additional Faculty Mentor Name

Joan Lin-Cereghino

Abstract/Artist Statement

The yeast Pichia pastoris is known to be efficient at expressing and producing recombinant proteins. Previous studies successfully produced the maltose binding protein (MBP), a type of "escort" protein that aids protein folding and purification. We expressed enhanced green fluorescent protein (EGFP) fused to either the N-terminus of MBP (MBP-EGFP, pJV4) or to the C-terminus MBP (EGFP-MBP, pVJ103). Surprisingly, MBP- EGFP was proteolyzed before secretion, but EGFP-MBP was secreted intact. The objective was to find out if the two fusions followed different paths in the cell by using fluorescence microscopy. This led to the development of a protocol for visualizing EGFP in Pichia pastoris cells. Our results suggest that depending on its position in the fusion, EGFP followed a different route in the cell.

Location

Grave Covell

Start Date

21-4-2012 10:00 AM

End Date

21-4-2012 12:00 PM

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Apr 21st, 10:00 AM Apr 21st, 12:00 PM

Visualizing the Pathways of MBP-EGFP Fusions with Fluorescence Microscopy

Grave Covell

The yeast Pichia pastoris is known to be efficient at expressing and producing recombinant proteins. Previous studies successfully produced the maltose binding protein (MBP), a type of "escort" protein that aids protein folding and purification. We expressed enhanced green fluorescent protein (EGFP) fused to either the N-terminus of MBP (MBP-EGFP, pJV4) or to the C-terminus MBP (EGFP-MBP, pVJ103). Surprisingly, MBP- EGFP was proteolyzed before secretion, but EGFP-MBP was secreted intact. The objective was to find out if the two fusions followed different paths in the cell by using fluorescence microscopy. This led to the development of a protocol for visualizing EGFP in Pichia pastoris cells. Our results suggest that depending on its position in the fusion, EGFP followed a different route in the cell.