Stocks, Christian F.2022-06-132022-06-132022https://hdl.handle.net/10092/103778http://dx.doi.org/10.26021/12877Genetic modification is vital for modern microbial research. In this thesis, genetic modification explored in bacteria with the use of plasmid vectors that contain modified transposons. Transformation of Gram-positive bacteria was explored through addition of the Himar1 transposon to the previously constructed chromatic bacteria toolbox. Conjugation was then used to insert the Himar-based plasmids into a range of Gram-positive and Gram-negative bacteria. Results of this showed that the Himar-based plasmids could not be used to produce insertion mutants of any Gram-positive strains. However, a handful of Gram-negative strains could be tagged using these plasmids including strains not tagged with the original chromatic bacteria toolbox. Following production of the Himar-based plasmids, transformation protocols were optimised in the Gram-positive strain Arthrobacter sp. Leaf145. Transformation allows for greater amounts of plasmid insertions than conjugation and would increase the number of insertion mutants produced the Himar-based plasmids. However, even after successful optimisation of transformation, insertion mutants were not observed with the Himar-based plasmids in Arthrobacter sp. Leaf145. During transformation optimisation, Tn1409- based plasmids were capable of producing insertion mutants in Arthrobacter sp. Leaf145. Therefore, the use of these plasmids was further explored for native protein over-expression in Arthrobacter sp. Leaf145. Along with genetic analysis of Arthrobacter sp. Leaf145, new Tn1409-based plasmids were constructed which allowed for expression of new genes in Arthrobacter sp. Leaf145. Due to time constraints, the plasmids that would allow for homologous protein over-expression in Arthrobacter sp. Leaf145 could not be tested. However, these are available for use in future research.enAll Rights ReservedTheses / Dissertations