title: [Construction of an activation tagging library of Arabidopsis and cloning for mutant genes].
abstract: Activation tagging is an important strategy in plant genomics by generating gain-of-function mutants. In this work, a library of Arabidopsis mutants was constructed by in planta transformation mediated by Agrobacterium tumefaciens containing an activation tagging vector pSKI015 with herbicide Basta as a selection marker (Fig. 1). Among 20000 independent transformants, 38 lines, i.e. about 0.2% of T(1) progeny, show visible morphological phenotypic variations (Fig. 2). Results of Southern blot analysis revealed that most of the transformants have more than three copies of T-DNA insertion (Fig. 3). Plasmid rescue and TAIL-PCR were used to recover the flanking genomic sequences of mutated target genes as the first step towards mutant gene cloning (Fig. 4, 5).
authors.affiliation: Institute of Vegetable Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Zhi-Miao
authors.initials: ZM
authors.lastname: Li
authors.forename: Hai-Kuo
authors.initials: HK
authors.lastname: Zhang
authors.forename: Jia-Shu
authors.initials: JS
authors.lastname: Cao
authors.forename: Zu-Hua
authors.initials: ZH
authors.lastname: He
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D009154:Mutation / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction