Induction of phenotypic variation by activation of genes harbouring a maize Spm element in their promoter regions using a TnpA-VP16 fusion protein.
For many crops, a narrowing genetic base is becoming an increasingly significant problem for improvements made through breeding. Commonly used breeding procedures systematically reduce genetic diversity within elite gene pools. Here we describe a new technique for activation of genes in lines carrying Spm or dSpm transposon insertions. Activation of genes in Arabidopsis harbouring Spm or dSpm insertions in their promoters can be induced by over-expression of the TnpA-VP16 fusion protein, which binds Spm ends and activates local transcription. As a result, a variety of phenotypes are recovered from multiple-copy Spm lines in Arabidopsis. Application of this technique to a number of Spm insertion collections in Arabidopsis provides a valuable approach for new insights into plant gene functions. It also provides a proof-of-principle demonstration that the method could be used to generate new variation in elite lines of maize.
Sorokin
Alexander P
AP
Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.
Walsh
Sean
S
Baumann
Kim
K
Nichols
John
J
Bevan
Michael
M
Jones
Jonathan D G
JD
Martin
Cathie
C
Clarke
Jonathan H
JH
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D015152:Blotting, Northern; D004251:DNA Transposable Elements / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D014644:Genetic Variation; D023281:Genomics / Q000379:methods; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D016254:Mutagenesis, Insertional; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D003313:Zea mays / Q000235:genetics*
D004251:DNA Transposable Elements; D017405:Herpes Simplex Virus Protein Vmw65; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins