An efficient cell-free translation system from Aspergillus nidulans and in vitro translocation of prepro-alpha-factor across Aspergillus microsomes.
We describe the preparation of an in vitro translation system from heat shock-treated Aspergillus nidulans, capable of supporting efficient and faithful synthesis of proteins from natural and in vitro transcribed eukaryotic messages. In vitro synthesized prepro-alpha-factor was translocated across Aspergillus nidulans microsomal membranes in either the homologous A. nidulans or a yeast cell-free system. The translocated prepro-alpha-factor was protected from digestion by protease and glycosylated to higher MW forms.
Devchand
M
M
Allelix Inc. Biopharmaceutical Division, Mississauga, Ontario, Canada.
Gwynne
D
D
Buxton
F P
FP
Davies
R W
RW
D001233:Aspergillus nidulans / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism*; D012331:RNA, Fungal / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins
D005656:Fungal Proteins; C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins