- Poster Presentation
- Open Access
The expression of truncated form of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) from genetically modified plant in Escherichia coli
https://doi.org/10.1186/1475-2859-5-S1-P83
© Stuchlík et al; licensee BioMed Central Ltd. 2006
- Published: 10 October 2006
Keywords
- Codon Usage
- Horizontal Gene Transfer
- Enzymatic Property
- Full Length Gene
- pET28a Plasmid
Background
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 [1]. The codon usage of both forms was originally optimized for plant [2]. 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.
Results
Physical maps of plasmids pKK1, pKK2, pMD2, pMD72.
Complementation test of the CP4 aro A phenotypes.
Plasmids in E. coli strain | 2% Glp | 3% Glp | 4% Glp | MM |
---|---|---|---|---|
pKK1 in SV1TcR | +++ | ++ | + | +++ |
pKK2 in SV1TcR | +++ | ++ | + | +++ |
pKK233-2 in SV1TcR | - | - | - | - |
pKK233-2 in JM106 | ND | ND | ND | +++ |
Growth curves of E. coli SV1TcR with plasmids pKK1 and/or pKK2 in M9 liquid medium. Both strains were grown in concentration of 0.5% glyphosate.
Conclusion
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.
Declarations
Acknowledgements
This work was supported by grant of Slovak Grant Agency APVT-20-17102.
Authors’ Affiliations
References
- Stuchlík S, Natarajan S, Turña J: Spontaneously generated truncated forms of CP4 EPSPS protein. FEBS Journal. 2005, 272: Supplement 1. Abstracts 30th FEBS Congress and 9th IUBMB Conference, Budapest, Hungary 2–7 July 2005Google Scholar
- Harrison LA, Bailey MR, Naylor MW, Ream JE, Hammond BG, Nida DL, Burnette BL, Nickson TE, Mitsky TA, Taylor ML, Fuchs RL, Padgette SR: The expressed protein in glyphosate-tolerant soybean, 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4, is rapidly digested in vitro and is not toxic to acutely gavaged mice. J Nutr. 1996, 126: 728-740.Google Scholar
Copyright
This article is published under license to BioMed Central Ltd.