Rose M D, Price B R, Fink G R
Mol Cell Biol. 1986 Oct;6(10):3490-7. doi: 10.1128/mcb.6.10.3490-3497.1986.
We have developed a protocol for efficient fusion of spheroplasts of the same mating type. Nuclear fusion in this whole-cell system is also efficient and closely parallels nuclear fusion in heterosexual mating of intact cells. In the spheroplast fusion system, nuclear fusion is dependent on both the KAR1 gene and prior exposure to alpha factor. The major products of nuclear fusion in the spheroplast fusion assay were true diploids that were homozygous at the mating-type locus. An additional 10% of the products were cells of ploidy greater than diploid. The dependence of nuclear fusion on alpha factor treatment could not be replaced by synchronization in G1 by mutations in CDC28 and CDC35 or by prior arrest in stationary phase. These data suggest that nuclear fusion is not a constitutive function of the nucleus, but rather is specifically induced by mating hormone.
我们已经开发出一种用于高效融合相同交配型原生质球的方案。在这个全细胞系统中的核融合也是高效的,并且与完整细胞的异性交配中的核融合非常相似。在原生质球融合系统中,核融合依赖于KAR1基因以及先前对α因子的暴露。原生质球融合试验中核融合的主要产物是在交配型位点纯合的真正二倍体。另外10%的产物是多倍体大于二倍体的细胞。核融合对α因子处理的依赖性不能通过CDC28和CDC35中的突变在G1期进行同步化或通过先前在稳定期停滞来替代。这些数据表明核融合不是细胞核的组成功能,而是由交配激素特异性诱导的。