ALBERT

Description:    Activator/Dissociation ( Ac/Ds ) transposable elements have been used in maize insertional mutagenesis as a complement to Mutator ( Mu ). In this study, to further improve the efficiency of the Ac / Ds mutagenesis system, we adopted apt1 - m1 (Ac) on the long arm of chromosome 9 (9L) as a donor Ac to create an Ac insertion library. This system is based on the negative selection pressure against the donor Ac , and it was highly efficient for isolating new transposition events. We obtained 9,625 transposition events from 1083 F1 ears with an average transposition rate of 8.66 % (rates ranged from 1.11 to 29.73 %). We also adopted a modified PCR-based genome walking strategy to improve the efficiency of the new method for isolating transposon-flanking sequences. This method is more efficient than the Southern-based method that was used in previous studies. A validation step was developed to distinguish transposon tags derived from newly transposed Ac or Ds elements. Using this PCR-based method, we isolated 67 inheritable flanking sequences from the apt1 - m1 (Ac) transposition library; of these, 51 were confirmed as tr- Ac -flanking sequences and 11 were tr- Ds -flanking sequences. Similar to other Ac donors from different loci, the apt1 - m1 (Ac) system also exhibited a preference for short distance transposition. In this study, we have further improved the Ac mutagenesis system in maize for gene isolation and functional genomics studies. Content Type Journal Article Pages 1-11 DOI 10.1007/s10709-012-9685-2 Authors Fei Wang, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Pengfei Li, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Yuanping Tang, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Jun Fan, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Dabin Xu, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Shengming Guo, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Zhengkai Xu, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Rentao Song, School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai, 200444 People’s Republic of China Journal Genetica Online ISSN 1573-6857 Print ISSN 0016-6707