Expression vector p301-bG1 contains a gus gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes (Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 μg/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.
PEG介导下香菇的转化
孙丽 许伟宏 蔡华清 胡鸢雷 林忠平*
（北京大学生命科学学院蛋白质工程和植物基因工程国家重点实验室，北京100871）
摘要：表达载体p301-bG1含有香菇(Lentinus edodes(Berk.)Sing)三磷酸甘油醛脱氢酶启动子驱动下的gus基因和除草剂抗性基因.利用PEG法实现了p301-bG1对香菇原生质体的转化.香菇原生质体与经PEG纯化的质粒DNA混合,用PEG处理后培养于含40μg/mL除草剂的CYM再生平板上,得到了抗除草剂和有GUS活性的转化菌株.虽然这种方法转化效率较低,但不需要昂贵的仪器和限制性内切酶,为蘑菇的分子育种研究提供了一种简便经济的转化方法.
关键词： 香菇；转化；三磷酸甘油醛脱氢酶基因启动子；gus；PEC
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