拟南芥两个MYB基因标签融合蛋白转基因植株的获得和鉴定

Formation of the Transgenic Arabidopsis Plant with Two MYB Genes Tagging the Fusion Protein and Its Identification

[目的]进行染色体免疫共沉淀(ChIP)试验,挖掘MYB类基因At3g11280和At5g05790所调控的下游基因。[方法]在构建了标签融合蛋白植物表达载体的基础上,利用农杆菌介导的转化将这些载体转入拟南芥植物中,获得转基因植株,并用Western blotting检测标签融合蛋白在转基因植株中的表达。[结果]拟南芥植株转基因的效率很高,4种标签蛋白融合载体的转化均获得了20株以上的转基因植株。随机对4种转基因植株的8株T1代植株进行Western blotting分析,结果表明4,种转基因植株中均鉴定出阳性植株。融合蛋白的条带大小在30 kD左右,将显色信号条带与Marker比对,显示条带大小与预测蛋白大小相符。[结论]这些有融合蛋白表达的转基因植株为ChIP试验的进行提供了重要材料。

[Objective] The downstream genes regulated by the MYB gene： At3g11280 and At5g05790,were developed through the experiment in chromosome immunoprecipitation（ChIP）.[Method] Based on the construction of the plant expression vectors tagging fusion protein,the transgenic Arabidopsis plants were produced after these vectors were introduced into the plants by means of the agrobacterium-mediated transformation,which expression in transgenic plants was detected by Western blotting method.[Result] The transgenic efficiency of Arabidopsis plants was very high and more than 20 transgenic plants were produced under the transformation of four-tagged protein fusion vectors.The Western blotting analysis result from four random 8 T1 plants of four types of transgenic plants showed that the positive plants could be founded in the four types of transgenic plants.The band of fusion protein was about 30 kD and it can be seen that the size of displaying band was consistent with that of predicted protein after the band with color signal was compared with the Marker.[Conclusion] These transgenic plants expressing fusion protein would be the important material for ChIP experiment.