Construction and application of efficient Ac-Ds transposon tagging vectors in rice.
Transposons are effective mutagens alternative to T-DNA for the generation of insertional mutants in many plant species including those whose transformation is inefficient. The current strategies of transposon tagging are usually slow and labor-intensive and yield low frequency of tagged lines. We have constructed a series of transposon tagging vectors based on three approaches: (i) AcTPase controlled by glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain (GVG) chemical-inducible expression system; (ii) deletion of AcTPase via Cre-lox site-specific recombination that was initially triggered by Ds excision; and (iii) suppression of early transposition events in transformed rice callus through a dual-functional hygromycin resistance gene in a novel Ds element (HPT-Ds). We tested these vectors in transgenic rice and characterized the transposition events. Our results showed that these vectors are useful resources for functional genomics of rice and other crop plants. The vectors are freely available for the community.
Qu
Shaohong
S
Department of Plant Pathology, Ohio State University, Columbus OH 43210, USA.
Jeon
Jong-Seong
JS
Ouwerkerk
Pieter B F
PB
Bellizzi
Maria
M
Leach
Jan
J
Ronald
Pamela
P
Wang
Guo-Liang
GL
D001483:Base Sequence; D015139:Blotting, Southern; D003433:Crosses, Genetic; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry; D003907:Dexamethasone / Q000494:pharmacology; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics*; D018745:Genome, Plant / Q000235:genetics; D019426:Integrases / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000235:genetics; D012275:Oryza / Q000187:drug effects / Q000235:genetics*; D030821:Plants, Genetically Modified; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics; D019895:Transposases / Q000378:metabolism
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D003907:Dexamethasone; C045073:Cre recombinase; D019426:Integrases; D019895:Transposases