Yamashiro C T, Yarden O, Yanofsky C
Department of Biological Sciences, Stanford University, CA 94305-5020.
Mol Gen Genet. 1992 Dec;236(1):121-4. doi: 10.1007/BF00279650.
When Neurospora crassa is transformed using a Neurospora gene as the selectable marker, the vegetatively stable transformants obtained cannot be used successfully in a cross because the selectable marker will be inactivated by the process of RIP (repeat-induced point mutation). Introduction of the acetamidase-encoding gene amdS of Aspergillus nidulans into N. crassa by transformation yielded transformants that would grow in minimal medium containing acetamide as a sole nitrogen source. In mitotically stable transformants containing a single copy of the amdS gene, the capacity to utilize acetamide as a sole nitrogen source was maintained in the progeny of a sexual cross. Therefore, the A. nidulans amdS gene is an appropriate dominant selectable marker for use in transformation analyses with N. crassa in which sexual crosses will be subsequently performed.
当使用粗糙脉孢菌基因作为选择标记对粗糙脉孢菌进行转化时，所获得的营养稳定型转化体不能成功用于杂交，因为选择标记会因RIP（重复诱导点突变）过程而失活。通过转化将构巢曲霉的乙酰胺酶编码基因amdS导入粗糙脉孢菌，产生了能在以乙酰胺作为唯一氮源的基本培养基中生长的转化体。在含有单拷贝amdS基因的有丝分裂稳定型转化体中，利用乙酰胺作为唯一氮源的能力在有性杂交的后代中得以维持。因此，构巢曲霉amdS基因是用于粗糙脉孢菌转化分析的合适显性选择标记，后续将在其中进行有性杂交。
