A studv on the structure and function of a fungal β-1, 6-glucanase

从粗脉孢子菌中克隆了内切β-1，6-葡聚糖酶基因Neg1。Neg1全长1443bp，编码463个氨基酸的成熟内切β-1，6-葡聚糖序列和17个氨基酸的信号肽。从大肠杆菌中纯化的重组蛋白(Neg1)具有β-1，6-D-葡聚糖酶活性。在酵母菌中没有发现同源基因。β-1，6-葡聚糖酶基因(Neg1)通过Repar诱导点突变(RIP)被破坏。基因中断菌株(R12-1)不具有β-1，6-葡聚糖酶活性。表型分析表明，副产物没有明显的形态变化，但蛋白质变性剂如十二烷基硫酸钠或十六烷基三甲基溴化铵抑制了菌丝的生长。

A gene (neg1) encoding endo-β-1,6-glucanase from Neurospora crassa was cloned. The putative neg1 was 1443-bp long and encoded a mature endo-β-1,6-glucanpse protein of 463 amino acids and signal peptide of 17 amino.acids. The purified recombinant protein (Neg1) obtained from Escherichia coli showed β-1,6-D-glucanase activity. No homologous genes are found in yeasts. and fungi.The β-1,6-glucanase gene (neg1) was disrupted by using repar-induced point mutation(RIP). The gene disruptant strain (R12-1) had no β-1,6-glucanase activity. The phenotype analyses showed thet the sisuptant had no apparent morphological changes, but protein denaturing agents, such as SDS or CTAB inhibited the mycerial growth.