Production of a polymer-forming fusion protein in Escerichia coli strain BL21.
In the course of studying [PSI(+)], a yeast prion, we found inadvertently that Escherichia coli strain BL21 overproducing a fusion protein, in which the prion-domain of Sup35 was connected to the C terminus of glutathione S-transferase, grew normally to the stationary phase and rapidly decreased in colony-forming ability thereafter. Evidence indicated that protein polymers consisting mainly of the fusion protein GST-Sup35NM (about 70% of the mass) and its N-terminal fragments were formed in extract prepared from the cells producing GST-Sup35NM. It was further found that cells of strain BL21 accumulated the protein polymers during prolonged cultivation. Based on these results, we contend that the initially observed defect in colony forming ability is the direct or indirect consequence of intracellular formation and accumulation of the protein polymers.
Ono
Bun-Ichiro
B
Department of Biotechnology, Faculty of Science and Engineering, Ritsumeikan University. ono@se.ritsumei.ac.jp
Kubota
Masashi
M
Kimiduka
Hiroko
H
Kawaminami
Hiroshi
H
Ueto
Takamitsu
T
Yokosawa
Shin
S
Iseda
Masako
M
Yamamoto
Yumiko
Y
Murakami
Yoshikazu
Y
Yokota
Sadaki
S
D001704:Biopolymers / Q000096:biosynthesis*; D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D001704:Biopolymers; D011993:Recombinant Fusion Proteins