Abstract
Development of an antibiotic-free plasmid selection system based on glycine auxotrophy for recombinant protein overproduction in Escherichia coli
Highlights
Antibiotics and antibiotics resistance genes have been traditionally used for the selection and maintenance of recombinant plasmids in hosts such as E. coli
A glycine-auxotrophic strain of E. coli M15 has been created using PCR products [3]. This strain contains an internal deletion of the glyA gene, which encodes for serine hydroxymethyl transferase (SHMT), an enzyme involved in the main glycine biosynthesis pathway in E. coli [4]
By using SOE-PCR [6], the glyA gene from E. coli K-12 was fused to the weak constitutive promoter P3
Summary
Antibiotics and antibiotics resistance genes have been traditionally used for the selection and maintenance of recombinant plasmids in hosts such as E. coli. Development of an antibiotic-free plasmid selection system based on glycine auxotrophy for recombinant protein overproduction in * Corresponding author from The 4th Recombinant Protein Production Meeting: a comparative view on host physiology Barcelona, Spain.
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