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发明公开
CN104131026A 利用抗萎锈灵基因carboxin为筛选标记的球孢白僵菌遗传转化方法 无效 - 驳回

利用抗萎锈灵基因carboxin为筛选标记的球孢白僵菌遗传转化方法
摘要：
本发明公开了一种利用抗萎锈灵基因carboxin为筛选标记的球孢白僵菌遗传转化方法，包括以下步骤：1)遗传转化载体的构建；2)球孢白僵菌原生质体的制备及转化；3)PEG介导的遗传转化体系建立；4)平均转化效率分析；5)PCR检测；6)荧光观察。本发明利用抗杀菌剂萎锈灵基因为筛选标记，成功进行球孢白僵菌遗传转化，转化效率达到26个转化子/μgDNA，拓宽了球孢白僵菌遗传转化筛选标记基因范围，解决bar基因存在的安全性问题。
摘要（英）：

The invention discloses a beauveria bassiana genetic transformation method by the utilization of anti-carboxin gene as a selection marker, comprising the following steps: 1) construction of a genetic transformation vector; 2) preparation and transformation of a beauveria bassiana protoplast; 3) establishment of a PED-mediated genetic transformation system; 4) analysis of average transformation efficiency; 5) PCR detection; and 6) fluorescence observation. By using the anti-carboxin gene as a selection marker, beauveria bassiana genetic transformation is successfully implemented. Transformation efficiency reaches 26 transformants/microgram DNA. The selection marker gene range for beauveria bassiana genetic transformation is widened, and safety problem of a bar gene is solved.

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