title: High-level expression of staphylococcal nuclease A in Escherichia coli.
abstract: The staphylococcal nuclease A gene has been successfully cloned and overexpressed in E. coli under the transcriptional control of the bacteriophage lambda PRPL promoters regulated by the temperature sensitive repressors. The SDS-PAGE analysis demonstrates that the nuclease A is produced to the extent of as much as 60% of the total cellular protein. The N-terminal analysis of the nuclease A shows that the amino terminal formyl methionine residue of the enzyme is precisely processed. The recombinant nuclease A with full activity is finally obtained after appropriate solubilization--denaturation and renaturation treatment. The conformational identity of the renatured nuclease A in different conditions is also studied by using hydrophobic interaction chromatography on a phenyl-superose HR5/5 column.
authors.affiliation: Institute of Biophysics, Chinese Academy of Sciences, Beijing.
authors.forename: G
authors.initials: G
authors.lastname: Jing
authors.forename: L
authors.initials: L
authors.lastname: Liu
authors.forename: Z
authors.initials: Z
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Zhou
authors.forename: Q
authors.initials: Q
authors.lastname: Zou
mesh_terms: D001483:Base Sequence; D015342:DNA Probes; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D008836:Micrococcal Nuclease / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics
chemical_list: D015342:DNA Probes; D008836:Micrococcal Nuclease