转座子Ac/Ds的水稻转化及杂交后代中Ds的跳跃分析

 摘要: 利用根癌农杆菌介导的转化法， 把双元表达载体CamDs（含有激活标签和基因捕获器结构）和含有玉米Ac转座酶的质粒(NeaAc)转入水稻。PCR结果表明Ac和Ds已整合到水稻的基因组中。转Ac植株与转Ds植株杂交，获得了12个杂交组合。杂交F1代水稻苗经过抗生素筛选，得到108株同时含有Ac和Ds因子的水稻苗。Basta抗性检测了Ds因子在杂交F1代中的跳跃情况，发现转座频率为13%。Ds空供体位点的PCR扩增结果与Basta抗性检测一致。另外，对跳动过的植株进行部分组织GUS染色表明Ds因子中的基因捕获器可以捕获到基因的表达。

Transformation of Ac/Ds into Rice and Ds Transposition Analysis of Hybrids

A binary vector CamDs carrying the maize transposon Ds with activation tagging and gene trap was constructed. The maize transposon Ac/Ds was transferred into rice (Oryza sativa subsp. japonica cv. Xiushui 11) by Agrobacterium-mediated transformation method. The integration of Ac/Ds into rice genome was confirmed by PCR. The Ds-inserted transgenic plants were crossed with the transgenic plants carrying Ac transposase and a population of 12 hybrids was obtained. One hundred and eight hybrids consisting of both Ds and Ac were obtained by resistance assaying. The result of the Basta resistance test indicated that the excision frequency of Ds element trans-activated by Ac transposase was 13%, PCR analysis showed the similar result. The GUS staining indicated that the gene trap system could capture the expression of the genes in rice genome.