In a systematic analysis of the adhesive and invasive capabilities of 1084 mutants, 10 mutants that showed more than a 50%% reduction in adhesion or invasion were obtained.
In the mutants that showed a significant decrease in the ability to adhere to HCT-8 cells, the Tail PCR method was used to identify the transposon insertion sites (Du et al. 2012).
Through screening of about 1 × 104 Escherichia coli BL21 DE3) transformants, we succeeded in isolating five mutants that showed a 35 217% increase in the secretion level of CGTase-HlyAs61 relative to the wild-type strain.
In the current study, to further our understanding of the molecular mechanisms involved in adhesion to and invasion into host cells in C. sakazakii, we constructed a Tn5 transposon mutant library and screened it to identify mutants that showed defects in adhesion or invasion.
The first biological clock genes, for example, were identified by examining Drosophila mutants that showed disrupted daily activity cycles.
Therefore, only mutants that showed a >50% decrease in current amplitude at 500 µM ACh were used for KB estimation.
To identify novel regulators of the hemolysins, we screened a transposon insertion library for mutants that showed altered toxin expression.
To identify mutants that showed altered hemolytic activity, we used a 48-well prong to transfer individual colonies onto sheep blood agar plates (SBA).
In this study, we screened a random transposon library for mutants that showed either an increase or decrease in hemolysin expression compared to wild type (WT).
