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

作　　者：郑卫红[1] 王瑜[2] 余增亮 

机构地区：[1]安徽省公安厅法医科 [2]中科院等离子体所离子束生物工程实验室,中国安徽合肥230031

出　　处：《激光生物学报》2000年第4期241-245,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金资助项目!( 1 9890 30 0 7);国家重点科技攻关项目资助课题!( 96- 538- 0 2- 0 1 )

摘　　要：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.

关 键 词：低能离子束 刻蚀 基因转移 烟草 外源基因转化 

分 类 号：S572.03[农业科学—烟草工业]