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用哈栖木霉ATCC48131的细胞溶壁酶从指状青霉的菌丝体中制备原生质体。将指状青霉ATCC10030于27℃培养在马铃薯右旋糖液体培养基中,静止培养24小时,菌丝用Whatman 1号滤纸过滤收集,然后用0.6％的氯化钠冲洗3次。原生质体是从0.8克分子甘露糖醇溶液(pH5.5～6.0)的菌丝悬液(50～100毫克/毫升)中,用5～7％(W/V)对哈栖木霉中得到的细胞壁溶解酶,在每分钟100转的往复式摇床上,30℃下,摇4小时浸提。采用二相梯度离心,从剩余菌丝体碎片中分离出原生质体。1毫升1M硫酸镁同1毫升粗制原生质悬
Protoplasts were prepared from the mycelium of Penicillium dicots using cell wall lysing enzymes of Trichoderma harzianum ATCC 48131. Penicillium albopictus ATCC10030 was cultivated in potato dextrose broth at 27 ° C and incubated for 24 hours. The mycelium was collected by filtration using Whatman filter paper 1 and then rinsed three times with 0.6% sodium chloride. Protoplasts were obtained from mycelial suspensions (50-100 mg / ml) of 0.8 molar mannitol solution (pH 5.5-6.0) with 5-7% (w / v) Of the cell wall lytic enzyme, in reciprocating shaker at 100 revolutions per minute, 30 ° C, shake 4 hours leaching. Protoplasts were isolated from remaining mycelial fragments using two-phase gradient centrifugation. 1 ml of 1 M magnesium sulfate and 1 ml of crude protoplasm suspension