Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 95, Issue 2, Pages 93-101Publisher
HUMANA PRESS INC
DOI: 10.1385/ABAB:95:2:093
Keywords
Escherichia coli; L-asparaginase; gene expression; plasmid stability; DNA sequence
Ask authors/readers for more resources
The L-asparaginase (ASN) from Escherichia coli AS1.357 was cloned as a DNA fragment generated using polymerase chain reaction technology and primers derived from conserved regions of published ASN gene sequences. Recombinant plasmid pASN containing ASN gene and expression vector pBV220 was transformed in different E. coli host strains. The activity and expression level of ASN in the engineering strains could reach 228 IU/mL of culture fluid and about 50% of the total soluble cell protein respectively, more than 40-fold the enzyme activity of the wild strain. The recombinant plasmid in E. coli AS1.357 remained stable after 72 h of cultivation and 5 h of heat induction without selective pressure. The ASN gene of E. coli AS1.357 was sequenced and had high homology compared to the reported data.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available