Deletion of HindIIR and HindIIIR improves DNA transfer via electroporation toHaemophilus influenzaeRd

Abstract

ABSTRACTHaemophilus influenzaeis a bacterial species of interest for its medical relevance and utility as a model system. Despite its role in several landmark molecular and synthetic biology studies,H. influenzaeremains underexplored as a potential chassis organism. The limited availability of reliable and convenient transformation methods and genetic tools forH. influenzaeare obstacles to this end. However, a strain ofH. influenzaeRd KW20 lacking the type II restriction endonucleases HindII and HindIII has previously been developed. Here, we show that this strain is more readily transformable by electroporation than wild-type Rd KW20. We also developed a series of multi-host plasmids carrying antibiotic selection and fluorescent visual markers based on the pSU20 vector. The availability ofH. influenzaeΔHindII/III, paired with the electroporation method and plasmids presented here, will promote the exploration ofH. influenzaeas a host organism for synthetic biology applications.