Department of Biological Sciences, Laboratory for Molecular Biology, University of Illinois at Chicago, 900 South Ashland Avenue, Chicago, IL 60607, USA.
Yeast. 2010 Mar;27(3):159-66. doi: 10.1002/yea.1740.
The yeast deletion library is a collection of over 5100 single gene deletions that has been widely used by the yeast community. The presence of a non-Mendelian element, such as a prion, within this library could affect the outcome of many large-scale genomic studies. We previously showed that the deletion library parent strain contained the [PIN(+)] prion. [PIN(+)] is the misfolded infectious prion form of the Rnq1 protein that displays distinct fluorescent foci in the presence of RNQ1-GFP and exists in different physical conformations, called variants. Here, we show that over 97% of the library deletion strains are [PIN(+)]. Of the 141 remaining strains that have completely (58) or partially (83) lost [PIN(+)], 139 deletions were able to efficiently maintain three different [PIN(+)] variants despite extensive growth and storage at 4 degrees C. One strain, cue2Delta, displayed an alteration in the RNQ1-GFP fluorescent shape, but the Rnq1p prion aggregate shows no biochemical differences from the wild-type. Only strains containing a deletion of either HSP104 or RNQ1 are unable to maintain [PIN(+)], indicating that 5153 non-essential genes are not required for [PIN(+)] propagation.
酵母缺失文库是一个由超过 5100 个单基因缺失株组成的集合,已被酵母界广泛应用。这个文库中存在非孟德尔元件,如朊病毒,可能会影响许多大规模基因组研究的结果。我们之前曾表明,缺失文库亲本菌株含有[PIN(+)]朊病毒。[PIN(+)]是 Rnq1 蛋白的错误折叠传染性朊病毒形式,在存在 RNQ1-GFP 的情况下会显示出独特的荧光焦点,并存在不同的物理构象,称为变体。在这里,我们表明,超过 97%的文库缺失株为[PIN(+)]。在 141 株仍保留[PIN(+)]的菌株中,完全(58 株)或部分(83 株)丢失[PIN(+)]的菌株,尽管在 4°C 下进行了广泛的生长和储存,仍能有效地维持三种不同的[PIN(+)]变体。一株 cue2Delta 显示出 RNQ1-GFP 荧光形状的改变,但 Rnq1p 朊病毒聚集体在生化上与野生型没有区别。只有含有 HSP104 或 RNQ1 缺失的菌株无法维持[PIN(+)],表明 5153 个非必需基因不需要用于[PIN(+)]的传播。