摘要
表达载体p3 0 1_bG1含有香菇 (Lentinusedodes (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p3 0 1_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 4 0 μg/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。
Expression vector p301-bG1 contains a Sw 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 mug/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.
