Chemical synthesis of a fully active transcriptional repressor protein.
Plasmid pLS1-encoded 45-amino acid transcriptional repressor CopG (formerly RepA) has been chemically synthesized. A one-step purification of the synthetic protein has been developed, which yields high levels of pure protein with low or no contamination of truncated products. We have compared some properties of the chemical CopG protein with those of the biologically purified CopG. The two proteins were indistinguishable in (i) their ability to generate specific protein-DNA complexes, (ii) their capacity to protect a restriction site included within the CopG DNA target, and (iii) in their in vitro capacity to specifically repress synthesis of copG mRNA.
del Solar
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G
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Cientificas, Madrid, Spain.
Albericio
F
F
Eritja
R
R
Espinosa
M
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000138:chemical synthesis* / Q000378:metabolism; D001483:Base Sequence; D004247:DNA; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D008969:Molecular Sequence Data; D011506:Proteins; D012097:Repressor Proteins / Q000138:chemical synthesis* / Q000378:metabolism; D015534:Trans-Activators; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D011506:Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004247:DNA; D004265:DNA Helicases