High-level expression of staphylococcal nuclease a in Escherichia coli
Jing, G.; Liu, L.; Liu, Z.; Zhou, B.; Zou, Q.
Chinese Journal of Biotechnology 10(1): 25-32
1994
ISSN/ISBN: 1042-749X PMID: 7993969 Document Number: 438223
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.