Complementation of myosin null mutants in Dictyostelium discoideum by direct functional selection.
Complementation of myosin null mutants in Dictyostelium discoideum by direct functional selection.
复制标题
通过直接功能选择来补充盘基网柄菌中的肌球蛋白无效突变体。
DOI：
10.1016/0012-1606(90)90260-p
复制
发表时间：
1990
影响因子：
2.7
通讯作者：
Spudich,JA
中科院分区：
文献类型：
--
作者：
Egelhoff,TT;Manstein,DJ;Spudich,JA
真核粘菌盘基网柄菌含有单一的传统肌球蛋白重链基因(MHCA)。该基因通过同源重组被删除的细胞系已有报道。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。我们在这里证明，克隆的mhcA基因可以通过使用直接功能选择重新导入这些细胞。这一选择是通过要求细胞能够在悬浮液中生长而强制进行的。由此产生的转化子看起来胞质分裂正常，也完全适合孢子发生，证实了重新导入肌球蛋白基因足以恢复这些特性。这些结果证明了一种拯救Dictyostelium突变的方法，该方法可能普遍适用于遗传产生的突变以及已设计的突变。
The eukaryotic slime moldDictyostelium discoideumcontains 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 clonedmhcA 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 inDictyosteliumwhich may be generally applicable for genetically created mutations as well as for mutations which have been engineered.