电击法介导的人参大片段DNA转化灵芝原生质体的研究  

Study on the Electroporation-mediated Transformation of Large DNA Fragment of Panax Ginseng into Protoplast of Ganoderma lucidum

作　　者：朱文静[1] 马飞[1] 孙春玉[1] 蒋世翠[1] 李珊珊[1] 张美萍[1] 王义[1] 

机构地区：[1]吉林农业大学生命科学学院,长春130118

出　　处：《特产研究》2015年第1期21-28,共8页Special Wild Economic Animal and Plant Research

基　　金：863项目(2013AA102604-3)

摘　　要：为探讨灵芝原生质体电击转化的最佳条件,以人参大片段DNA(80kb)为材料,通过电击转化法,将其转入6d菌龄的灵芝原生质体中。使用2mm电击杯的最佳转化条件为平均外加电场861.34V、大片段DNA∶原生质体为1∶4;PCR检测结果显示,载体左区699bp处和右区397bp处电泳检测结果均显示有目的条带,初步判断人参大片段DNA已经转入灵芝原生质体中;镜检发现,转化子的菌丝分枝少,仍然存在锁状联合,菌丝体边缘更致密、整齐且生长速度更快,灵芝酸含量增高。This study used the large fragment of 80kb DNA from Panax g/nseng as experimental materials to find out the optimum conditions ofElectroporation- mediated transformation. The most suitable age of C, anoderma luc/dum for the transformation were 6d; The best voltage was861.34V when the electrode spacing was 2mm;The most suitable proportion between large fragments and protoplast was l：4;The expectedbands which were consistent with the left（699bp） and right（397bp） flanking sequences were found, which preliminarily proved that the largefragment DNA had been transformed into Ganoderma luc/dum ;The mycelia of positive transformation still had the clamp - connection and lessbranches than the original ones. The mycelia of the transformats grew faster and the margin was denser. The ganoderic acid contents were in-

关 键 词：大片段DNA 模式真菌 电击转化法 灵芝原生质体 

分 类 号：S567.51[农业科学—中草药栽培]