Abstract
A fusion gene encoding ribonuclease T1-EGFP (rntA-egfp) was constructed and expressed to use it as a tool for studies on the secretory pathway in Aspergillus oryzae. The successful secretion of the intact RntA-EGFP fusion protein was detected by fluorescence measurement and Western analysis. With use of the RntA-EGFP system, we were able to see high fluorescence at hyphal tips and observe concentrated fluorescence at septa in basal cells during growth at optimal conditions. Cold or heat shock during growth caused the accumulation of EGFP fluorescence in vacuoles.
MeSH terms
-
Aspergillus oryzae / genetics
-
Aspergillus oryzae / growth & development
-
Aspergillus oryzae / metabolism
-
Aspergillus oryzae / physiology*
-
Blotting, Western
-
Cold Temperature
-
Coumarins / chemistry
-
Green Fluorescent Proteins
-
Hyphae / metabolism
-
Luminescent Proteins / biosynthesis
-
Luminescent Proteins / chemistry
-
Luminescent Proteins / genetics*
-
Luminescent Proteins / metabolism
-
Plasmids / genetics
-
Recombinant Fusion Proteins / analysis*
-
Recombinant Fusion Proteins / biosynthesis
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / metabolism*
-
Ribonuclease T1 / biosynthesis
-
Ribonuclease T1 / genetics*
-
Ribonuclease T1 / metabolism
-
Spectrometry, Fluorescence
-
Vacuoles / chemistry
-
Vacuoles / ultrastructure
Substances
-
7-amino-4-chloromethylcoumarin
-
Coumarins
-
Luminescent Proteins
-
Recombinant Fusion Proteins
-
Green Fluorescent Proteins
-
Ribonuclease T1