泡桐丛枝植原体染色体全长及两个rRNA操纵子定位研究

The chromosome size and the locations of the two rRNA operons of paulownia witches'-broom phytoplasma

本实验以泡桐丛枝植原体为材料,运用差速离心和脉冲场电泳(Pulsed Field Gel Electrophoresis,PFGE),获得了无寄主DNA污染、完整的泡桐丛枝植原体染色体,并用Southern blot杂交进行了验证,建立了植原体全基因组的研究方法。同时对泡桐丛枝植原体染色体进行内切酶I-CeuI不完全酶切,测定了染色体全长,并定位了2个非连锁的rRNA操纵子在染色体上的位置,完成了泡桐丛枝植原体部分物理图谱的构建。实验结果表明,泡桐丛枝植原体染色体全长约为1 100kb,2个rRNA操纵子在染色体上相距500～600kb。

We established a method for research of the paulownia witches＇-broom phytoplasma（PaWB） genome using differential centrifugation and pulsed field gel electrophoresis（PFGE）.And the method confirmed by Southern blot.Partial digestions with I-CeuI and PFGE were used to determine the size of the PaWB chromosome,which was estimated to be about 1100kb.And we also located the two unlinked rRNA operons which were 500~600kb apart on PaWB chromosome.