A pink bacterium as a reporter system signaling expression of a recombinant protein.
We describe a set of expression vectors, pEX-PINK/0-3, for high-level production of (un)fused target proteins. The vectors incorporate a 'pink' reporter element, which signals in vivo the expression status of a target gene. A target sequence is cloned between the lambda PL promoter and the downstream mammalian cytochrome b5 gene. Thermo-induction drives transcription of a dicistronic mRNA from which the target protein and cytochrome b5 are independently and concurrently synthesized. Positive expression is indicated by visual transformation of bacteria from a grey/translucent to a bright pink color derived from tandemly expressed holocytochrome b5. The signal can be monitored in vivo spectrophotometrically.
Kaderbhai
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N
Department of Biochemistry, University of Wales, Aberystwyth, UK.
Gallagher
J
J
He
M
M
Kaderbhai
M A
MA
D001483:Base Sequence; D003001:Cloning, Molecular; D003124:Colorimetry; D015786:Cytochromes b5 / Q000235:genetics / Q000378:metabolism; D004247:DNA; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics* / Q000378:metabolism; D013057:Spectrum Analysis; D014908:Triticum / Q000201:enzymology / Q000235:genetics*
D011994:Recombinant Proteins; D004247:DNA; D015786:Cytochromes b5; D012273:Ribulose-Bisphosphate Carboxylase