Cloning ospA of Borrelia Burgderferi for Plant Expression

Abstract

Abstract Borrelia burgdorferi (Lyme disease) is a vector-borne disease that is both very common in the Mid-Atlantic region and which causes long term negative health effects. It is primarily spread to humans by deer ticks, and because one of the main ways that it is passed between ticks is by previously infected mouse blood, our goal is to design an oral vaccine which decreases infection rates in mice. Our goal is to do this by inserting outer surface protein A (ospA) of B. burgdorferi into a plant vector: pCambia 1201. Agrobacterium tumefaciens was used as a means of introducing the vector (bearing B. burgdorferi ospA) into Arabidopsis thaliana. The pCambia vector was introduced into A. tumefaciens competent cells after being purified from E. coli colonies which had undergone a similar transformation process, as E. coli is more suitable for cloning. The successfully electroporated A. tumefaciens was then plated and colonies were picked and grown as liquid cultures. Those liquid cultures transformed with both the ospA bearing pCambia and a control pCambia were used for floral dip of Arabidopsis thaliana (both ospA bearing and pCambia only control plants). These dipped plants’ seeds have been tested for successful transformation, and one plant has been found to contain the sequence for ospA. Continued floral dips will be undertaken to increase the number of positive plants, and to develop a proper control for testing efficacy as a vaccine. Any additional seeds of positive plants will be planted and crossed to ensure use of only plants which express pCambia in later testing, and to ensure that the process has been successful. Supported by Frostburg State University Department of Biology and FSU Foundation