Mu-lac insertion-directed mutagenesis in a pectate lyase gene of Erwinia chrysanthemi.
The pelC gene, which encodes one of the five major pectate lyase (PL) isoenzymes in Erwinia chrysanthemi 3937, designated PLc, was subcloned from a hybrid lambda phage into a pBR322 derivative and mutagenized with a mini-Mu-lacZ transposable element able to form fusions to the lacZ gene. One plasmid (pAD1) which had an inactivated pelC gene and a Lac+ phenotype was selected in Escherichia coli. This plasmid was introduced into Erwinia chrysanthemi, and the pelC::mini-Mu insertion was substituted for the chromosomal allele by homologous recombination. This strain lacks the PLc isoenzyme. This Erwinia chrysanthemi strain has a Lac+ phenotype that is inducible by polygalacturonate, as are the wild-type PL activities.
Diolez
A
A
Coleno
A
A
D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004885:Erwinia / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011133:Polysaccharide-Lyases / Q000235:genetics*
D004251:DNA Transposable Elements; D004262:DNA Restriction Enzymes; D011133:Polysaccharide-Lyases; C023911:pectate lyase