Fluorescent protein fusions for protein localization in plants.
Protein localization in living plant cells is commonly studied using fluorescent protein fusions. Stable transformation of plant cells requires the use of binary vectors, which are larger and not as amenable to genetic manipulation as animal cell transfection vectors. Binary vectors containing fluorescent protein fusion constructs are prepared using standard molecular biological techniques. Fusion genes as well as promoters and selection markers are stably incorporated into the plant cell genome via Agrobacterium-mediated transfer. Presented here are a series of protocols that detail binary vector construction, bacterial transformation, and a rapid transient assay technique that can be used to evaluate fusion protein fluorescence in leaves.
Runions
John
J
Research School of Life Sciences, Oxford Brookes University, Oxford, UK.
Hawes
Chis
C
Kurup
Smita
S
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D018744:DNA, Plant / Q000737:chemistry / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D059828:Plant Cells; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D010944:Plants / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012231:Rhizobium / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D018744:DNA, Plant; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins