低能离子束介导外源基因转化烟草的研究

Studies on the Transferring Gene to Tobacoo by the Low Energy Ion Beam

以烟草 NC— 89种子为材料 ,用显微扫描电镜 ( ESM)和电子自旋共振 ( ESR)波谱仪研究氮离子束对烟草种子表面的刻蚀作用及能量沉积产生自由基的间接效应 ,为离子束介导转移外源基因提供了形态结构依据。将烟草种子用 2 0 Kev的氮离子束处理后 ,浸入含有 PB 1 2 1质粒的缓冲介质中 ,在含有卡那霉素1 0 0 mg/ L的 MS0 培养基上继代筛选 ,得到 3株抗性植株。取抗性植株的叶片 ,经组织培养后得到再生抗性植株。经过 PCR及 southern杂交分析 。

The etching function of ion beam on the tobacoo seeds(NC-89) was investigated by means scanning electron microscope(SEM) and elecron spin resonance(SER), which provides the morphological structure basis of using ion beam in transgenic technique. The seeds of tobacoo were treated by 20Kev N + ion beam, then immediately immerged in SSC solution by containing PB II21 DNA (50mg/ml) which encodes NPT II R and GUS gene. Three NPT II R plants were obtained by selecting in two months tissue culture on MS media supplemented with 100mg/L kanamycin. Leaves from NPT II R plants were differentiated and regenerated into NPT II R plants. PCR and Southern blot results showed that the NPT II R and GUS gene on the plasmid PB II21 had been integrated into the genome of the tobacco.