Plasmid to generate Mycobacteria mutants

Evolutionary thrift : mycobacteria repurpose plasmid diversity during adaptation of type

Mycobacteria have a distinct secretion system, termed type VII (T7SS), which is encoded by paralogous chromosomal loci (ESX) and associated with pathogenesis, conjugation, and metal homeostasis. Evolution of paralogous gene families is of interest because duplication is an important mechanism by which novel genes evolve, but there are potential conflicts between adaptive forces that stabilize d...

Temperature-sensitive mutants of the Streptomyces plasmid pIJ702.

DNA from the Streptomyces plasmid pIJ702 was mutagenized in vitro using hydroxylamine and transformed into Streptomyces lividans. One plasmid with temperature-sensitive replication (pMT660) and one plasmid with a temperature-sensitive tyrosinase (pMT661) were isolated. The plasmid pMT661 contains a novel PstI restriction endonuclease site within the tyrosinase gene.

Isolation and characterization of novel plasmid-encoded umuC mutants.

Most inducible mutagenesis in Escherichia coli is dependent upon the activity of the UmuDC proteins. The role of UmuC in this process is poorly understood, possibly because of the limited number of genetically characterized umuC mutants. To better understand the function of the UmuC protein in mutagenic DNA repair, we have isolated several novel plasmid-encoded umuC mutants. A multicopy plasmid...

Generation of large deletion mutants from plasmid DNA.

BENCHMARKS In the course of our work on the vitamin D receptor (VDR) protein, we became interested in the generation of several large deletion mutant versions of this protein for structure-function relationship studies. One goal was the deletion of a 50 amino acid sequence from the 427 amino acids of the native protein in addition to the introduction of several point mutations throughout the re...

Plasmid-instability in acrA mutants of Escherichia coli K12.