HIGHLY-EFFICIENT TRANSFORMATION OF THE HOMOBASIDIOMYCETE SCHIZOPHYLLUM-COMMUNE TO PHLEOMYCIN RESISTANCE
HIGHLY-EFFICIENT TRANSFORMATION OF THE HOMOBASIDIOMYCETE SCHIZOPHYLLUM-COMMUNE TO PHLEOMYCIN RESISTANCE
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DOI：
10.1007/bf00313808
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发表时间：
1994-08-01
期刊：
影响因子：
2.5
通讯作者：
WESSELS, JGH
中科院分区：
文献类型：
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作者：
SCHUREN, FHJ;WESSELS, JGH
将同源裂殖酵母的甘油醛-3-磷酸脱氢酶(GPD)基因的调控序列与新链霉菌的ble基因的编码序列进行了融合，该基因编码一个金霉素结合蛋白。当在0.5M的硫酸镁中进行再生时，所得到的构建物以较高的频率(每10(7)个原生质体中高达10(4)个转化子/每10(7)个原生质体中的DNA)将链霉菌转化为对金霉素抗性。一个与来自尼杜拉曲霉的调控序列类似的构建未能给出转化子，这表明了同源调控序列对公社曲霉基因表达的重要性。同源的GPD启动子可缺失至-130位，而不影响转化子的数量。这是第一个使用抗生素耐药性的同源担子菌的有效稳定转化系统。
Regulatory sequences of the glyceraldehyde-3-phosphate-dehydrogenase (GPD) gene from the homobasidiomycete Schizophyllum commune were fused to the coding sequence of the ble gene from Streptoalloteichus hindustanus, which codes for a phleomycin-binding protein. The resulting construct transformed S. commune to phleomycin resistance at a high frequency (up to 10(4) transformants/mu g DNA per 10(7) protoplasts) when regeneration was done in 0.5 M MgSO4. A similar construct with regulatory sequences from Aspergillus nidulans failed to give transformants, showing the importance of homologous regulatory sequences for the expression of genes in S. commune. The homologous GPD promoter could be deleted up to position -130 without any effect on the number of phleomycin-resistant transformants. This is the first effective stable transformation system in a homobasidiomycete employing antibiotic resistance.