Preparation and use of recombinant plasmids containing genes for alkaline phosphatases

US4375514A · kind A · utility

Assignee

• SCHERING CORPORATION · US

Inventors

• Gerhard Siewert · Berlin, DE
• Werner Boidol · Berlin, DE
• Joachim Daum · Berlin, DE

Key dates

• Filing date: Aug 1, 1980
• Grant date: Mar 1, 1983
• Expiry date: Aug 1, 2000

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S435/881
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

PhoA mutant E. coli SB44, prepared by mutation of E. coli HB101 with an N-nitroso compound, can be used to identify and isolate recombinant plasmids into which a phoA gene has been incorporated. These plasmids can be used to transform bacteria which can be cloned and incubated to provide alkaline phosphatase in high yield. Moreover, these plasmid vectors can be modified in various ways so that the N-terminal amino acid sequence of phoA is followed in reading phase by the DNA coding for some other protein. In turn, these new plasmids can be used to transform bacteria which can be cloned and incubated to produce fusion proteins comprising the desired other protein in high yield and outside of the cell membrane in the periplasmatic space.