Chen C Y, Oppermann H, Hitzeman R A
Nucleic Acids Res. 1984 Dec 11;12(23):8951-70. doi: 10.1093/nar/12.23.8951.
DNA sequences normally flanking the highly expressed yeast 3-phosphoglycerate kinase (PGK) gene have been placed adjacent to heterologous mammalian genes on high copy number plasmid vectors and used for expression experiments in yeast. For many genes thus far expressed with this system, expression has been 15-50 times lower than the expression of the natural homologous PGK gene on the same plasmid. We have extensively investigated this dramatic difference and have found that in most cases it is directly proportional to the steady-state levels of mRNAs. We demonstrate this phenomenon and suggest possible causes for this effect on mRNA levels.
通常位于高表达的酵母3-磷酸甘油酸激酶(PGK)基因两侧的DNA序列,已被置于高拷贝数质粒载体上的异源哺乳动物基因旁,并用于酵母中的表达实验。对于迄今为止用该系统表达的许多基因,其表达水平比同一质粒上天然同源PGK基因的表达水平低15至50倍。我们广泛研究了这一显著差异,发现在大多数情况下,它与mRNA的稳态水平成正比。我们证明了这一现象,并提出了影响mRNA水平的可能原因。