PMID:1761034
| Citation |
Nikkila, H, Gennis, RB and Sligar, SG (1991) Cloning and expression of the gene encoding the soluble cytochrome b562 of Escherichia coli. Eur. J. Biochem. 202:309-13 |
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| Abstract |
The gene for the soluble cytochrome b562 from Escherichia coli B has been cloned on a SalI fragment. The analysis of the gene reveals the presence of a leader sequence in front of the sequence encoding the mature protein. Expression of cytochrome b562 using the lac-promoter produced the protein to a level of 3-5% of total protein. This over-production enables employment of a simple, high-yield purification protocol to obtain homogeneous cytochrome b562. Spectroscopic and N-terminal sequence analyses of the purified protein demonstrate that it is identical to the chromosomally expressed cytochrome b562 purified and characterized from E. coli B [Itagaki, E. & Hager, L.P. (1966) J. Biol. Chem. 241, 3687-3695]. It is demonstrated that the genomic sequence codes for a classic N-terminal signal sequence and that mature cytochrome b562 is translocated to the periplasmic space. |
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| Keywords |
Amino Acid Sequence; Base Sequence; Blotting, Southern; Chromosomes, Bacterial; Cloning, Molecular; Cytochrome b Group/genetics; Cytochrome b Group/isolation & purification; DNA, Bacterial/genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins; Gene Expression; Genes, Bacterial; Molecular Sequence Data; Plasmids |
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