用植株转化法将GUS基因导入桑树幼苗的研究

Transducting GUS Gene Into Mulberry Seedling by Plant Transformation Technique

为探讨简便易行的农杆菌介导的转化方法 ,用工程农杆菌A 110株 (LBA4 4 0 4 /pIG12 1 Hm)对桑幼苗子叶腋隙进行刺伤感染 ,感染部位长出的桑芽表型异常。取其基因组DNA进行分子生物学鉴定 ,结果显示 :以基因组DNA为模板 ,经PCR反应后得到特异性扩增片段用SalⅠ酶解 ,酶切产物与预计大小完全相符 ;用GUS基因作探针进行Southern杂交和用Km基因作探针对基因组DNA进行Southern杂交 ,均获得较强的杂交信号。

In order to get convenient manipulative translation technique with Agrobacterium tumefaciens, the cotyledon leaf gap of mulberry seedlings was infected with Agrobacterium tumefaciens A-110 strain (LBA4404/pIG121-Hm) by perforation. It was observed that the mulberry buds on the infected position were abnormal, its genomic DNA was identified by molecular biology method. The results indicated as: The expected fragment was amplified by PCR using the genomic DNA as template, which was digested by Sal Ⅰ, and the fragments were similar to that we expected in size. The PCR product was hybridized with the genomic DNA by Southern bloting using the GUS gene or Km gene as probe, a strong signal was detected.