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Generation of an Escherichia coli lysA targeted deletion mutant by double cross‐over recombination for potential use in a bacterial growth‐based lysine assay
Author(s) -
Li X.,
Ricke S.C.
Publication year - 2003
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2003.01425.x
Subject(s) - escherichia coli , homologous recombination , mutant , auxotrophy , biology , dna , lysine , gene , strain (injury) , microbiology and biotechnology , mutation , genetics , chemistry , amino acid , anatomy
Aims: To generate a stable Escherichia coli lysine auxotroph for the lysine bioavailability assay. Methods and 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, Significance and Impact of the Study: The Δ lysA :: bla strain exhibited a linear response to lysine supplementation and can be used for quantifying lysine.