Egelhoff T T, Manstein D J, Spudich J A
Department of Cell Biology, Stanford University School of Medicine, California 94305.
Dev Biol. 1990 Feb;137(2):359-67. doi: 10.1016/0012-1606(90)90260-p.
The eukaryotic slime mold Dictyostelium discoideum contains a single conventional myosin heavy chain gene (mhcA). Cell lines in which this gene was deleted via homologous recombination have been previously reported. These myosin null cells were shown to be defective for cytokinesis and for sporogenesis. We demonstrate here that the cloned mhcA gene can be reintroduced into these cells by the use of a direct functional selection. This selection was imposed by demanding that cells be capable of growth in suspension. The resulting transformants appear normal for cytokinesis, and also are fully competent for sporogenesis, confirming that reintroduction of the myosin gene is sufficient to restore these properties. These results demonstrate a method for rescuing mutants in Dictyostelium which may be generally applicable for genetically created mutations as well as for mutations which have been engineered.
真核黏菌盘基网柄菌含有一个单一的传统肌球蛋白重链基因（mhcA）。先前已有报道通过同源重组删除该基因的细胞系。这些肌球蛋白缺失细胞在胞质分裂和孢子形成方面存在缺陷。我们在此证明，通过直接功能选择，克隆的mhcA基因可以重新导入这些细胞。这种选择是通过要求细胞能够在悬浮状态下生长来施加的。所得转化体在胞质分裂方面看起来正常，并且在孢子形成方面也完全具备能力，证实肌球蛋白基因的重新导入足以恢复这些特性。这些结果证明了一种拯救盘基网柄菌突变体的方法，该方法可能普遍适用于基因产生的突变以及经过工程改造的突变。
