A rapid procedure for cloning genes from lambda libraries by complementation of E. coli defective mutants: application to the fabE region of the E. coli chromosome.
I describe a general and rapid procedure allowing the isolation of specialized lambda transducing phages from a lambda library by lysogenic complementation of defective mutants of Escherichia coli. As an example, the cloning of the E. coli fabE gene and of two other adjacent genetic determinants is presented. Subcloning and determination of its nucleotide sequence reveals that fabE codes for the biotin carboxyl carrier protein (BCCP), one of the three subunits of acetyl coenzyme A carboxylase.
Alix
J H
JH
Institut de Biologie Physico-Chimique, University of Paris, France.
D000103:Acetyl-CoA Carboxylase / Q000235:genetics*; D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015723:Gene Library; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008025:Ligases / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014161:Transduction, Genetic
D008025:Ligases; D000103:Acetyl-CoA Carboxylase