TY - JOUR
T1 - Expression of the GFP-mammalian pleckstrin homology (PH) domain of the phospholipase C δ1 in Saccharomyces cerevisiae BY4741
AU - Perrine-Walker, Francine
AU - Payne, Jennifer
N1 - Funding Information:
J.P. was financially supported by the Australian Postgraduate Award. No funding was received to assist with the preparation of this manuscript.
Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature B.V.
PY - 2022/5
Y1 - 2022/5
N2 - Background: Pleckstrin homology (PH) domains are common modules of ∼120 amino acids found in proteins involved in signalling, cytoskeletal organization, membrane transport, and modification of phospholipids. Previous live cell studies have involved the use of the green-fluorescent protein (GFP) labelling of PH-domain of phospholipase C δ1 (PLC δ1) to study the interactions of molecules at the membrane interface. Methods and results: For this study, the aim was to construct and express the GFP-PH domain of PLC δ1 in the Saccharomyces cerevisiae BY4741. The transformants expressing GFP-PH domain of PLC δ1 displayed localised fluorescence to the cell periphery (plasma membrane) while the negative control expressed GFP within the cytoplasm only. No GFP was observed in the non-transformed yeast cells. Conclusions: Thus, this technique could be useful in future molecular interactions studies targeted specifically at the yeast cell membrane interface in live yeast cells.
AB - Background: Pleckstrin homology (PH) domains are common modules of ∼120 amino acids found in proteins involved in signalling, cytoskeletal organization, membrane transport, and modification of phospholipids. Previous live cell studies have involved the use of the green-fluorescent protein (GFP) labelling of PH-domain of phospholipase C δ1 (PLC δ1) to study the interactions of molecules at the membrane interface. Methods and results: For this study, the aim was to construct and express the GFP-PH domain of PLC δ1 in the Saccharomyces cerevisiae BY4741. The transformants expressing GFP-PH domain of PLC δ1 displayed localised fluorescence to the cell periphery (plasma membrane) while the negative control expressed GFP within the cytoplasm only. No GFP was observed in the non-transformed yeast cells. Conclusions: Thus, this technique could be useful in future molecular interactions studies targeted specifically at the yeast cell membrane interface in live yeast cells.
KW - Confocal microscopy
KW - Green fluorescent protein
KW - Yeast
UR - https://www.scopus.com/pages/publications/85129573971
U2 - 10.1007/s11033-022-07414-0
DO - 10.1007/s11033-022-07414-0
M3 - Article
C2 - 35526246
AN - SCOPUS:85129573971
SN - 0301-4851
VL - 49
SP - 4123
EP - 4128
JO - Molecular Biology Reports
JF - Molecular Biology Reports
IS - 5
ER -