Muhammad Ehsaan

TL;DR: The improved ClosTron system supersedes the prototype plasmid pMTL007 and the original method, and exploits the potential of Group II introns more fully, and means mutants can now be constructed with very little time and effort for the researcher.

TL;DR: Novel DNA integration strategies are explored that exploit activation or inactivation of genes leading to a selectable phenotype, and asymmetrical regions of homology to control the order of recombination events to open the way to reliable integration of DNA including large synthetic constructs in diverse microorganisms.

TL;DR: An efficient procedure for making precise alterations to the C. difficile genome by pyrE-based allelic exchange is developed, allowing complementation studies to be undertaken at an appropriate gene dosage, as opposed to the use of multicopy autonomous plasmids.

TL;DR: A simple roadmap is formulated whereby the necessary gene systems maybe developed and deployed and the creation of a pyrE mutant (a uracil auxotroph), initially aided by ClosTron technology, but ultimately made using a special form of allelic exchange termed ACE (Allele-Coupled Exchange).

TL;DR: Enc encouraging data suggest that the novel enzyme and strain engineering approach represent a promising platform for the clinical development of CDEPT, and a novel nitroreductase from Neisseria meningitidis, NmeNTR, which is able to activate CB1954 at clinically-achievable serum concentrations is identified.