Generation of an Escherichia coli lysA targeted deletion mutant by double cross-over recombination for potential use in a bacterial growth-based lysine assay.
OBJECTIVE
To generate a stable Escherichia coli lysine auxotroph for the lysine bioavailability assay.


RESULTS
An E. coli lysine auxotrophic strain was constructed by deleting the entire lysA gene and replacing it with a gene that confers resistance to ampicillin (bla). The linear DNA contained 50 bp homologous sequence of upstream of lysA in one end and 50 bp of downstream of lysA in the other end.


CONCLUSIONS
The deltalysA::bla strain exhibited a linear response to lysine supplementation and can be used for quantifying lysine.
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Poultry Science Department, Texas A&M University, Kleberg Center, TX 77843-2472, USA.
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D000668:Ampicillin Resistance / Q000235:genetics; D001426:Bacterial Proteins; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D017353:Gene Deletion; D005798:Genes, Bacterial; D008239:Lysine / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D002262:Carboxy-Lyases; C079912:LysA protein, Bacteria; C020031:diaminopimelic acid decarboxylase; D008239:Lysine