Team:UCSF/Data

Our control for this lab was the EBY100 yeast strain of S. cerevisiae. Cultures of EBY100 (some transformed with GFP) were grown in liquid cultures of S Raffinose media and S Raffinose/1% Galactose overnight and examined the next day on the microscope. From our observations under the microscope, EBY100 rarely aggregate into clusters, while occasionally forming clumps of several cells.

Cadherin

Since cadherins only form dimers under the presence of calcium ions, we were able to use that variable as a second control mechanism. We observed that EBY100 transformed with cadherin would not form clumps until calcium was added to the cultures. However, when EDTA was added to the cultures, the clusters seen previously had dispersed.

HWP1

We did not have a clear understanding of how HWP1 really bind in Candida albicans and help form biofilms. So at first we just simply grew HWP1 in liquid cultures and we saw enormous clusters. However, we were unsure if the clumping was really caused by random adhesion between HWP1 or just adhesion to the budding from an EBY100 cell expressing HWP1. We then mixed EBY100 expressing HWP1 and EBY100 expressing GFP and HWP1 to see if the clusters were actually formed by cells randomly adhering to one and other. And from our observations, we were able to conclude that the clumping was indeed random. We also mixed normal EBY100 with EBY100 transformed with HWP1 and GFP to test if HWP1 was adhering just to the surface of EBY100, but we haven’t tested this hypothesis enough to reach a decisive conclusion.

Mgfp5