Development of a reporter peptide that catalytically produces a fluorescent signal through α-complementation.
In α-complementation, inactive N-terminal (α-domain) and C-terminal (ω-domain) fragments of β-galactosidase associate to reconstitute the active protein. To date, the effect of α-domain size on α-complementation activity has not been systematically investigated. In this study, we compared the complementation activities of α-domains of various sizes using an in vitro system. We found that the complementation activities are similar for α-domains comprising between 45 and 229 N-terminal residues but are significantly decreased for those containing less than 37 residues. However, these smaller α-domains (15 and 25 residues) exhibited sufficient α-complementation activity for application as reporters.
Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Kotaro
K
Nishiyama
Norikazu
N
Ichihashi
Yasuaki
Y
Kazuta
Tetsuya
T
Yomo
D000595:Amino Acid Sequence / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics
D010455:Peptides; D001616:beta-Galactosidase
cell-free translation system; reporter gene; reporter peptide; α-complementation; β-galactosidase