Jin Wei-zheng, Wang Shao-min, Xu Min, Duan Rui-jun, Wu Ping
The State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310029, China.
J Zhejiang Univ Sci. 2004 Apr;5(4):390-9. doi: 10.1631/jzus.2004.0390.
Insertion mutagenesis has become one of the most popular methods for gene functions analysis. Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice. The excision of Ds element was examined by PCR amplification. The excision frequency of Ds element varied from 0% to 40% among 20 F(2) populations derived from 11 different Ds parents. Southern blot analysis revealed that more than 70% of excised Ds elements reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice. The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots, flowers or seeds. The GUS positive lines will be useful for identifying gene function in rice.
插入诱变已成为基因功能分析最常用的方法之一。在此,我们报道了一种用于水稻基因标签的双元件Ac/Ds转座子系统,该系统包含增强子捕获和基因捕获。通过PCR扩增检测Ds元件的切除情况。在来自11个不同Ds亲本的20个F(2)群体中,Ds元件的切除频率在0%至40%之间变化。Southern杂交分析表明,超过70%的切除Ds元件重新插入水稻基因组,且重新插入的Ds元件中有70%以上位于水稻染色体的不同位置。组织化学GUS分析结果表明,28%的增强子捕获标签植株和22%的基因捕获标签植株在叶片、根、花或种子中表现出GUS活性。GUS阳性株系将有助于鉴定水稻中的基因功能。