This chapter discusses epitope tagging for the detection of fusion protein expression in transgenic plants. The chapter describes the development of the c-myc epitope tag system for the detection of transprotein expression in transgenic tobacco plants. An expression vector for plant c-myc epitope tagging was constructed and introduced into tobacco plants via Agrobacterium-mediated transformation. The results show that c-myc epitope tagged trans-protein can be detected in extracts from as little as 5 mg of leaf tissue. The Mycl-9El0 monoclonal antibody shows no cross-reactivity to endogenous tobacco or Arabidopsis thaliana proteins. In addition, the antibody can be used to localize expression in various tissues using the tissue print method. The ability of the Mycl-9E10 antibody to detect the 10-amino acid c-myc epitope on Westerns blots and tissue prints and the absence of antibody cross reactivity to endogenous tobacco and Arabidopsis proteins make this epitope–antibody pair an excellent choice as a general marker for protein expression in transgenic plants.
All Science Journal Classification (ASJC) codes
- Cell Biology