Emergence of transmissible antibiotic resistance (AbR) in Enterobacteriaceae is a major problem worldwide. The most important vectors of this transmissible AbR are conjugative plasmids those acquires very quickly and, once acquired, becomes fixed in the bacterial accessory genome by ‘addiction systems’ that poison cells from which the AbR plasmid is lost. Our aim is to develop ‘therapeutic plasmids’ which ultimately will eradicate addictive AbR plasmids from microbiome and restore normal microflora. Here we examined in vitro curing of two locally endemic plasmids, pEl1573 (IncL/M, with pemIK only) encodes gentamicin resistance and AbR to almost all available β-lactams (with the carbapenemase gene blaIMP-4) and pJIE512b (IncI1, with pndBCA) resistance to cephalosporin antibiotics and β-lactamase inhibitors (due to AmpC β-lactamase gene blaCMY-2). These plasmids are both 100% retained after 100 successive passages without antibiotic selection in nutrient broth. We constructed pJIMK24, 25 by cloning various rep and antitoxin genes into the small high-copy chloramphenicol resistant pBC-SK vector and specific ‘therapeutic plasmids’ by replacing the resistance region and toxin genes from our two example conjugative resistance plasmids pEl1573 and pJIE512b with tetA or fosA3 to create pJIMK46 and pJIMK56, retaining only antitoxin genes and introducing resistance to tetracycline and fosfomycin. These ‘therapeutic plasmids’ were then introduced into different Enterobacteriaceae species carrying pEl1573 or pJIE512b by transformation or mating. 100% of tested Enterobacteriaceae species were cured from target AbR plasmids by introduction with therapeutic plasmids. Mouse gut was colonised with target AbR plasmid (either pEl1573 or pJIE512b) and specific therapeutic plasmid (pJIMK46 or pJIMK56) was then administered with gelatine food for three days. Complete cure of target AbR plasmid has been achieved from the mice gut by therapeutic plasmid treatment. Therapeutic plasmids were then lost spontaneously from mice gut within few days. This approach offers hope of not only rendering resistant pathogen susceptible to therapy but also of a cure for antimicrobial resistance and potentially restoring normal microflora.