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. 1993 Apr 1;291(Pt 1):315–322. doi: 10.1042/bj2910315

The expression of Escherichia coli threonine synthase and the production of threonine from homoserine in mouse 3T3 cells.

W D Rees 1, S M Hay 1
PMCID: PMC1132519  PMID: 8385938

Abstract

We have subcloned the coding sequence for the Escherichia coli threonine synthase gene into a eukaryotic expression vector based on the simian-virus-40 early promoter. When mouse 3T3 cells which already expressed homoserine kinase were transfected with the new plasmid, the cells were able to incorporate radioactivity from [14C]homoserine into their cell proteins. Stable cell lines were established by co-transfecting 3T3 cells with the plasmid coding for threonine synthase and another coding for homoserine kinase and G-418 (Geneticin) resistance. Cells were selected for G-418 resistance and then screened for an ability to synthesize threonine from homoserine and incorporate it into the cell protein. A cell line which expressed both the homoserine kinase and threonine synthase genes was capable of growth in a threonine-deficient medium containing homoserine.

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Selected References

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