Murai N, Li Z J, Kawagoe Y, Hayashimoto A
Department of Plant Pathology and Crop Physiology, College of Agriculture, Louisiana State University, Baton Rouge 70803-1720.
Nucleic Acids Res. 1991 Feb 11;19(3):617-22. doi: 10.1093/nar/19.3.617.
Transposition of the maize Activator (Ac) element was observed in transgenic rice. After protoplast transformation, Ac excision from an interrupted hygromycin phosphotransferase gene was monitored by appearance of the hygromycin-resistant colonies. The frequency of Ac excision, based on the biological assay was up to 19%. Southern hybridization analysis indicated that at least one copy per genome of the hygromycin-resistance gene was reconstituted after Ac excision and that the transposed Ac element was reintegrated into the rice genome. Analysis of DNA sequences at 14 empty donor sites indicated that the Ac element was excised in rice in a similar manner as maize. The excision of an Ac mutant in which a 1.3 kbp Tn903 fragment was inserted at a unique BamHI site so as to disrupt binding of the putative transposase was not detected by DNA analysis. These results demonstrated that the maize Ac element might be used as an effective heterologous transposon for mutagenesis and gene tagging in rice, an important food crops.
在转基因水稻中观察到玉米激活子(Ac)元件的转座。原生质体转化后,通过潮霉素抗性菌落的出现来监测Ac从中断的潮霉素磷酸转移酶基因上的切除。基于生物学测定,Ac切除频率高达19%。Southern杂交分析表明,Ac切除后每个基因组至少有一个潮霉素抗性基因拷贝被重建,并且转座的Ac元件重新整合到水稻基因组中。对14个空供体位点的DNA序列分析表明,Ac元件在水稻中的切除方式与玉米相似。通过DNA分析未检测到在唯一的BamHI位点插入1.3 kbp Tn903片段以破坏假定转座酶结合的Ac突变体的切除。这些结果表明,玉米Ac元件可能用作水稻(一种重要粮食作物)诱变和基因标签的有效异源转座子。