Methods for generation and analysis of fluorescent protein-tagged maize lines.
The use of fluorescent proteins to localize gene products in living cells has revolutionized cell biology. Although maize has excellent genetics resources, the use of fluorescent proteins in maize cell biology has not been well developed. To date, protein localization in this species has mostly been performed using immunolocalization with specific antibodies, when available, or by overexpression of fluorescent protein fusions. Localization of tagged proteins using native regulatory elements has the advantage that it is less likely to generate artifactual results, and also reports tissue-specific expression patterns for the gene of interest. Fluorescent protein tags can also be used for other applications, such as protein-protein interaction studies and purification of protein complexes. This chapter describes methods to generate and characterize fluorescent protein-tagged maize lines driven by their native regulatory elements.
DuPont Knowledge Centre, Hyderabad, India.
Amitabh
A
Mohanty
Yan
Y
Yang
Anding
A
Luo
Anne W
AW
Sylvester
David
D
Jackson
D016960:Agrobacterium tumefaciens / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D005821:Genetic Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D050436:Regulatory Elements, Transcriptional; D003313:Zea mays / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D000097573:Red Fluorescent Protein
D001426:Bacterial Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C054562:yellow fluorescent protein, Bacteria