Ahmad Ausaf, Kabir M Anaul, Kravets Anatoliy, Andaluz Encarnación, Larriba German, Rustchenko Elena
Department of Biochemistry and Biophysics, Box 712, University of Rochester Medical Center, Rochester, NY 14642, USA.
Yeast. 2008 Jun;25(6):433-48. doi: 10.1002/yea.1597.
Electrophoretic karyotyping of the Candida albicans revealed a different migration pattern of ChR in three different stocks of the sequencing strain SC5314. In one stock, the high instability of ChR size prevented the migration of ChR as a compact band; ChR appeared, instead, as a smear. In some stocks, ChR and/or Ch1 ploidy diminished, suggesting mixed populations of disomic and monosomic cells. Similarly, some stocks of widely used derivatives CAI4 and BWP17 contained smearing of ChR. In addition, the most manipulated strain in the lineage of SC5314, the last derivative, BWP17, acquired an increase in the size of Ch7b and revealed an unusual property. BWP17 did not tolerate a well-established procedure of telomere-mediated fragmentation of a chromosome; the remaining intact homologue always duplicated. We suggest that some stocks of SC5314 are unstable and that BWP17 may not be appropriate for general studies. Instead of BWP17 or CAI4, we recommend using for general research CAF4-2, which is a relatively stable Ura- derivative, and which has been successfully used for more than a decade in our laboratory.
白色念珠菌的电泳核型分析显示,在测序菌株SC5314的三种不同菌株中,染色体区域(ChR)呈现出不同的迁移模式。在一种菌株中,ChR大小的高度不稳定性阻止了其以紧密条带的形式迁移;相反,ChR呈现为一片拖尾。在一些菌株中,ChR和/或Ch1倍性降低,表明存在二体和单体细胞的混合群体。同样,一些广泛使用的衍生物CAI4和BWP17的菌株也存在ChR拖尾现象。此外,在SC5314谱系中经过最多操作的最后一个衍生物BWP17菌株,其Ch7b大小增加并表现出一种不寻常的特性。BWP17不能耐受一种成熟的染色体端粒介导的断裂程序;剩余完整的同源染色体总是会复制。我们认为SC5314的一些菌株不稳定,并且BWP17可能不适用于一般研究。我们建议在一般研究中使用CAF4-2,而不是BWP17或CAI4,CAF4-2是一种相对稳定的尿嘧啶营养缺陷型衍生物,并且在我们实验室已经成功使用了十多年。