Biochemistry, Biophysics, and Structural Biology Commons^™

Transposon Mutagenesis In Mycobacterium Avium Subspecies Paratuberculosis, John P. Bannantine, Denise K. Zinniel, Raul G. Barletta

School of Veterinary and Biomedical Sciences: Faculty Publications

While transposon mutagenesis has been developed for Mycobacterium avium subspecies paratuberculosis (Map), relatively few laboratories have adopted this important genetic tool to examine gene function and essentiality. Here we describe the construction of a Map transposon library using the Himar1 mariner transposon, but concepts can also be applied to the Tn5367 transposon, which has also been used by our group. Delivery of the transposon is by a temperature-sensitive phagemid, ϕMycoMarT7, and plating transductants requires patience and specialized media due to length of incubation required to observe colonies. Several transposon mutants obtained from these libraries have been tested in vaccine and …