University of Nottingham

Morvus Technology Limited

Nigel Peter Minton

John Timothy Heap

Ying Zhang

John Heap

Katrin Schwarz

The present application discloses a DNA molecule comprising a modified Group II intron which does not express the intron-encoded reverse transcriptase but which contains a modified selectable marker gene in the reverse orientation, wherein the marker gene comprises a Group I intron in forward orientation of causing expression in a bacteria cell of the class Clostridia and wherein the DNA molecule comprises sequences that allow for the insertion of the RNA transcript of the Group II intron in the chromosome of a bacterial cell of the class Clostridia. A method of introducing a nucleic acid molecule into a site of a DNA molecule in a bacterial cell of the class Clostridia is also provided. The DNA molecule and the method are useful for making mutationsspp.

DNA molecules and methods

The present invention provides a method of treatment for cancer in a subject comprising the administration of a prodrug, wherein the subject is colonised with an obligate anaerobic microorganism capable of expressing a polypeptide having nitroreductase activity, wherein the polypeptide exhibits preferential reduction of CB1954 to a 4-hydroxylamine (4HX) derivative instead of a 2-hydroxylamine derivative; and is capable of reducing CB1954 to the 4HX derivative with a Kof less than 30 micromolar. The present invention further provides an isolated polypeptide, nucleic acids encoding said polypeptide, vectors and host cells comprising said nucleic acids and vectors and uses of the above in treating solid tumours.

Treatment for cancer

The present invention now provides a conditional vector comprising DNA encoding for: (i) an inducible expression cassette comprising an inducible promoter operably linked to a plasmid replication region; and (ii) a selectable marker.

Conditional vectors and uses thereof

A method of double crossover homologous recombination in a host cell comprising: a first homologous recombination event between a donor DNA molecule comprising a first element of a selectable allele and an acceptor DNA molecule comprising a second element of the selectable allele in the host cell, thereby to form a product of the first homologous recombination event in the host cell; and a second homologous recombination event within the product of the first homologous recombination event, thereby to form a product of the second homologous recombination event in the host cell which confers a selectable phenotype on the host cell, wherein the selectable phenotype arises following and in dependency on the formation of a selectable allele from the first and second elements of the selectable allele.

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Nottingham, United Kingdom

Nigel Peter Minton