双向选择标记amdS在黑曲霉中自发突变及amdS-检测  

Spontaneous Mutation of the Bi-directional Selection Marker amdS Gene and Screening amdS-Strains in A.niger

作　　者：赵颖[1] 巴再华[1] 张春艳[1] 

机构地区：[1]济宁医学院

出　　处：《济宁医学院学报》2008年第4期283-285,共3页Journal of Jining Medical University

摘　　要：目的将传统上作为营养缺陷标记的amdS进行诱变,使其成为双向选择标记,有利于构建黑曲霉多基因缺失菌株。方法以已有amdS作选择标记的黑曲霉双基因缺失株为出发突变株,在以尿素为唯一氮源的氟代乙酰氨培养基上筛选amdS-突变菌株。结果将另一以amdS作选择标记的基因进行转化,得到一株三基因缺失菌株。结论在黑曲霉中利用amdS可作为双向选择标记的特点,构建多基因缺失菌株是行之有效的。Objective To construct the multiple gene deleted in A. niger, we let the amdS gene which traditionally is the auxotrophy selective marker to be mutated as the bi - directional selective marker. Methods A. niger△dppV△dppIV#2 -3 as the parental strain which has used the amdS gene as the selective marker can be grown on fluoridacetamide minimal medium plate （urea as the only nitrogenous matter）. Because only the strains whose amdS gene is spontaneous destroyed can better grow on the fluoridacetamide minimal medium plate, we screen amdS - strains from possible mutants. Results Then confirmed one strain F△dppV△dppIV#4.55 to be the recipient of mnn9 disruptant linetype gene segment which also uses amdS gene as the selective marker. The transformant △dppV△dppIV△mnn9#31 -7 was identified to be a dppIV,dppV,mnn9 triple disruption strain. Conclusion Using amdS gene as the bi -directional selective marker to construct the multiple genes deleted in A. niger is valid.

关 键 词：选择标记 突变 多基因缺失 

分 类 号：R197.32[医药卫生—卫生事业管理] R285.5[医药卫生—公共卫生与预防医学]