High impact information on fliG

• These were seen in wild-type cells and in a fliN mutant, but not in flhC or fliG mutants [1].
• The Mot- mutations of fliG isolated previously by random mutagenesis do not alter the putative active-site residues, but render the proteins abnormally susceptible to proteolysis, suggesting significantly altered conformations or reduced stabilities [2].
• Isolation of these mutants was facilitated by a "sensitizing" mutation (E232G) outside of the hinge region that was accidentally introduced during cloning of the chromosomal fliG gene into our vector plasmid [3].
• Class II suppressors were point mutations in the chromosomal motB or fliG genes [4].
• We have generated a class of cheY mutations selected for dominant suppression of fliG mutations [5].

Biological context of fliG

• DNA encoding FliG was mutagenized by error-prone amplification with Taq polymerase, mutant fliG genes were cloned (as DNA-binding domain-fliG gene fusions) in S. cerevisiae by gap repair of plasmid DNA, and mutants exhibiting an interaction defect were isolated in a two-hybrid screen [6].

Analytical, diagnostic and therapeutic context of fliG

• Engineering and amplifying fliG by PCR allowed convenient cloning into an efficient expression vector [7].
• Polyclonal antibodies (Ab) were generated against purified FliG and used in quantitative Western blots to determine that the wt expression level of fliG results in about 3700 FliG copies per cell [7].

References