New tool for metabolic pathway engineering in Escherichia coli: one-step method to modulate expression of chromosomal genes.
A simple and highly efficient method was developed to produce a library of Escherichia coli clones that express a particular chromosomal gene at a wide range of expression levels. The basic strategy was to replace all or part of the upstream region of a coding sequence containing the elements involved in its expression (promoter, operator, gene coding for a regulator, ribosome binding site, and start codon) with a PCR-generated library of expression cassettes.
Meynial-Salles
Isabelle
I
CRITT-Bioindustries CRT, INSA, DGBA, Toulouse, France.
Cervin
Marguerite A
MA
Soucaille
Philippe
P
D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic
D029968:Escherichia coli Proteins