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对高雄山虫草无性型细脚拟青霉的酶解条件、原生质体制备、电泳条件和核型进行了研究。以10mg/mL相同浓度的溶壁酶、纤维素酶和蜗牛酶配成混合酶解液,在30℃、120r/min振荡处理18h菌龄的菌丝2.5h,获得约107个/mL的原生质体悬浮液。浓缩至约108个/mL原生质体悬浮液与1.4%的低熔点琼脂糖等体积混合制胶,50℃蛋白酶K酶解48h,ET液洗3次(每次1h),浸没,4℃保存。以温汉逊氏酵母Hansenula wingei YB-4662-VIA和粟酒裂殖酵母Schizosaccharomyces pombe 972h-染色体DNA作为标准,利用钳位均匀电场脉冲电泳系统电泳,细脚拟青霉染色体组被分离成8条带,利用AlphaEase Fc软件分析,估计其大小在0.43-5.78Mb之间,大小分别为:0.43、2.45、2.55、2.88、3.28、3.94、4.70、5.78Mb,推测细脚拟青霉核型大小约为26.01Mb。
The enzymatic hydrolysis conditions, protoplast preparation, electrophoresis conditions and karyotype of Paecilomyces variotii from Kaohsiung were studied. 10 mg / mL of the same concentration of lywallzyme, cellulase and snail enzyme dubbed mixed enzyme solution, 30r, 120r / min shaken 18h strain mycelium 2.5h, get about 107 / mL protoplasm Body suspension. Concentrated to about 108 / mL protoplast suspension and 1.4% low melting point agarose and other volume mixing glue, 50 ° C protease K digestion 48h, ET liquid wash 3 times (each time 1h), immersion, 4 ℃ preservation. Using the Hansenula wingei YB-4662-VIA and Schizosaccharomyces pombe 972h-chromosomal DNA as a standard, the genome of P. minutus was separated into eight bands Band, using AlphaEase Fc software analysis, the estimated size of 0.43-5.78Mb, respectively, the size of: 0.43,2.45,2.55,2.88,3.28,3.94,4.70,5.78Mb, speculated that the foot of Penicillium karyotype size 26.01Mb.