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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。
The expression vector p3 0 1_bG1 contains the gus gene and the herbicide resistance gene driven by the glyceraldehyde-3-phosphate dehydrogenase promoter of Lentinusedodes (Berk.) Sing. The transformation of protoplast of mushroom was realized by PEG method. Mushrooms protoplasts were mixed with PEG purified plasmid DNA, treated with PEG and cultivated on CYM regeneration plates containing 40 μg / mL herbicide to obtain herbicide-tolerant and transformed strains with GUS activity. Although this method is less efficient, it does not require expensive instruments and restriction enzymes, which provides a simple and economical method for the molecular breeding of mushrooms.