Characterization of an Ac transposon system based on apt1-m1 (Ac) on the long arm of maize chromosome 9
Characterization of an Ac transposon system based on apt1-m1 (Ac) on the long arm of maize chromosome 9
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玉米 9 号染色体长臂上基于 apt1-m1 (Ac) 的 Ac 转座子系统的表征
DOI：
10.1007/s10709-012-9685-2
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发表时间：
2012-09
期刊：
影响因子：
1.5
通讯作者：
王飞
中科院分区：
文献类型：
--
作者：
王飞
激活子/解离(Ac/DS)转座元件已用于玉米插入突变，作为突变体(Mu)的补充。本研究采用9号染色体长臂上的APT1-M1(Ac)作为供体Ac，构建Ac插入文库，进一步提高Ac/ds突变体系的效率。该系统基于对供体Ac的负选择压力，对于分离新的转座事件是高效的。1083只F1耳共获得9625个易位事件，平均易位率为8.66%(1.11~29.73%)。我们还采用了一种改进的基于PCR的基因组行走策略，以提高新方法分离转座子侧翼序列的效率。这种方法比以前研究中使用的基于Southern的方法更有效。开发了一种验证步骤来区分来自新转座的Ac或DS元件的转座子标签。利用这种基于PCR的方法，我们从APT1-M1(Ac)转座文库中分离到67个可遗传的侧翼序列，其中51个被确认为tr-ac-侧翼序列，11个被确认为tr-ds-侧翼序列。与来自不同基因座的其他Ac供体类似，APT1-M1(Ac)系统也表现出对短距离转座的偏好。本研究进一步完善了玉米Ac诱变体系，用于基因分离和功能基因组学研究。
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.
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DOI：
10.1007/978-3-540-68922-5_11
发表时间：
2009
期刊：
--
影响因子：
--
作者：
A. Settles
通讯作者：
A. Settles
影响因子：
3.7
作者：
Ji J;Braam J
通讯作者：
Braam J
影响因子：
3.3
作者：
Zhennan Xu;H. Dooner
通讯作者：
Zhennan Xu;H. Dooner
DOI：
10.1128/9781555817954
发表时间：
2002
期刊：
--
影响因子：
--
作者：
N. Craig;M. Chandler;M. Gellert;A. Lambowitz;P. Rice;S. Sandmeyer
通讯作者：
N. Craig;M. Chandler;M. Gellert;A. Lambowitz;P. Rice;S. Sandmeyer
影响因子：
3.3
作者：
Conrad, Liza J.;Bai, Ling;Brutnell, Thomas P.
通讯作者：
Brutnell, Thomas P.