The expression of truncated form of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) from genetically modified plant in Escherichia coli
© Stuchlík et al; licensee BioMed Central Ltd. 2006
- Published: 10 October 2006
- Codon Usage
- Horizontal Gene Transfer
- Enzymatic Property
- Full Length Gene
- pET28a Plasmid
During the study of horizontal gene transfer of the aro A CP4 gene encoding CP4 EPSPS from genetically modified feed through gastrointestinal tract to bacteria living in animal gut we have observed beside full length gene also functional truncated one present in bacteria . The codon usage of both forms was originally optimized for plant . Therefore we have used two different E. coli expression systems (with tac promoter and T7 promoter) to compare enzymatic properties and functional activities of both forms of CP4 EPSPS protein.
Complementation test of the CP4 aro A phenotypes.
Plasmids in E. coli strain
pKK1 in SV1TcR
pKK2 in SV1TcR
pKK233-2 in SV1TcR
pKK233-2 in JM106
We can conclude from our results that truncated form of CP4 EPSPS (shorter to 31 amino acid residues on N-terminal end) can confer full function of native protein and these findings also should be taken into account in risk assessment of possible HGT from GM food/feed. The enzymatic properties and comparison of both EPSPS forms will be subject of further studies.
This work was supported by grant of Slovak Grant Agency APVT-20-17102.
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