Cross F R
Rockefeller University, New York, NY 10021, USA.
Yeast. 1997 Jun 15;13(7):647-53. doi: 10.1002/(SICI)1097-0061(19970615)13:7<647::AID-YEA115>3.0.CO;2-#.
One-step gene disruption constructs for disruption of HIS3, LEU2, TRP1 or URA3 with each of the other three markers have been constructed. All of these constructs have been tested and found to effectively convert markers either in gene disruptions or on plasmids. The 'swapped' strains allow the unambiguous genetic analysis of synthetic phenotypes with multiple genes, even if the original gene disruptions were made with the same marker. They also allow introduction of multiple plasmids in a single transformant, even if the original plasmids had the same marker, and allow transformation of plasmids into strains containing gene disruptions made with the same marker that is on the plasmids. These 'marker-swap' plasmids therefore eliminate the need for much subcloning to change markers. Marker-swapped alleles are acceptably stable mitotically and meiotically for most applications.
已经构建了一步基因破坏构建体,用于用其他三个标记中的每一个破坏HIS3、LEU2、TRP1或URA3。所有这些构建体都经过测试,发现在基因破坏或质粒中能有效转换标记。“交换”菌株允许对具有多个基因的合成表型进行明确的遗传分析,即使最初的基因破坏是用相同的标记进行的。它们还允许在单个转化体中引入多个质粒,即使原始质粒具有相同的标记,并且允许将质粒转化到含有与质粒上相同标记进行基因破坏的菌株中。因此,这些“标记交换”质粒消除了为改变标记而进行大量亚克隆的需要。对于大多数应用而言,标记交换等位基因在有丝分裂和减数分裂中具有可接受的稳定性。