Toxicity - pcDNA3 - crypticinitiation - (May/23/2001 )

Does anyone have any experience of toxicity problems / cryptic initiation when using pcDNA3 for subcloning in e.coli?

To date - we have experienced deletion problems in a number of host strains - all advice gratefully taken

I also faced the same problem of deletion in an attempt to clone a 900 bp fragment in a preexisting clone inpcDNA3 which would have given a total plasmid length.whenever we have tried to put large inserts into pcDNA3 the plasmids gets internal deletions.we have tried with bacterial hosts like DH5-alpha,TG-1 etc with the same result.