Biology Reference
In-Depth Information
CHAPTER
15
Live-Cell Imaging
of Microtubules
and Microtubule-Associated
Proteins in Arabidopsis
thaliana
Jessica Lucas
Department of Biology, Santa Clara University, Santa Clara, California, USA
CHAPTER OUTLINE
Introduction ............................................................................................................ 232
15.1 Protocols........................................................................................................234
15.1.1 Building Transgenic A. Thaliana .................................................. 234
15.1.1.1 Transgene Construction ..................................................... 235
15.1.1.2 Plant Transformation ......................................................... 236
15.1.1.3 Transgenic Plant Recovery................................................. 237
15.1.1.4 Screening T2 Seed Lines ................................................... 238
15.1.2 Sample Preparation .................................................................... 239
15.1.3 Imaging ..................................................................................... 240
15.2 Materials........................................................................................................242
15.2.1 Sterile Media for Sterile Plant Growth .......................................... 242
15.2.2 A. Tumefaciens Suspension for Plant Transformation .................... 242
15.2.2.1 Virulence Buffer................................................................. 242
Concluding Comments ............................................................................................. 243
Acknowledgment..................................................................................................... 243
References ............................................................................................................. 243
Abstract
Microtubules and microtubule-associated proteins (MAPs) play fundamental roles in
plant growth and morphogenesis. The ability to observe microtubules and MAPs in
living cells using fluorescent protein fusions has propelled plant scientists forward
and given them the opportunity to answer longstanding biological questions. In com-
bination with the genetic resources available in the model plant
Arabidopsis
