THE DEVELOPMENT OF A HETEROLOGOUS TRANSFORMATION SYSTEM FOR THE CELLULOLYTIC FUNGUS TRICHODERMA-REESEI BASED ON A PYRG-NEGATIVE MUTANT STRAIN
THE DEVELOPMENT OF A HETEROLOGOUS TRANSFORMATION SYSTEM FOR THE CELLULOLYTIC FUNGUS TRICHODERMA-REESEI BASED ON A PYRG-NEGATIVE MUTANT STRAIN
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DOI：
10.1007/bf00321118
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发表时间：
1990-07-01
期刊：
影响因子：
2.5
通讯作者：
DEGRAAFF, LH
中科院分区：
文献类型：
--
作者：
GRUBER, F;VISSER, J;DEGRAAFF, LH
通过对5-氟乳清酸的抗性筛选，从里氏木霉（Trichoderma reesei）QM 9414菌株中分离到6株尿苷营养缺陷型突变株，其中1株经放射性酶分析鉴定为外膜蛋白脱羧酶阴性（pyr-）。用粗糙脉孢菌的pyr 4基因（高达1500个转化体/μ g）和尼日尔曲霉的皮拉（700-800个转化体/μ g）实现向尿苷原养型的转化。在许多转化体中，PYR+功能似乎以染色体外DNA的形式存在。有证据表明转化体的稳定性与载体在基因组中的整合之间存在相关性，而当质粒发生自主复制时，获得不稳定的转化体。
Six uridine auxotroph mutants of Trichoderma reesei QM 9414 were isolated by resistance to 5-fluoroorotic acid and one strain was identified as OMP-decarboxylase negative (pyr-) by a radiometric enzyme assay. Transformation to uridine prototrophy was achieved with the pyr4 gene of Neurospora crassa (up to 1500 transformants/.mu.g) and with pyrA of Aspergillus niger (700-800 transformants/.mu.g). In many transformants the PYR+ function seems to be present as extrachromosomal DNA. There is evidence for a correlation between the stability of transformants and integration of the vector in the genome whereas unstable transformants are obtained when autonomous replication of the plasmid occurs.