Nasr F, Bécam A M, Brown S C, De Nay D, Slonimski P P, Herbert C J
Centre de Génétique Moléculaire, Laboratoire propre du CNRS, associé à l'Université Pierre et Marie Curie, Gif-sur-Yvette, France.
Mol Gen Genet. 1995 Nov 1;249(1):51-7. doi: 10.1007/BF00290235.
YBR1012 (YBR136w) is an essential gene from Saccharomyces cerevisiae identified during the systematic sequencing of part of the right arm of chromosome II. We previously constructed a conditional allele of YBR1012 based on antisense RNA, by inserting a small fragment of this gene downstream from the inducible UASGAL10-CYC1 promoter. Several other antisense RNA constructions have since been made and their activity tested. The response of the system appears to be very delicate, as the presence or absence of 13 nucleotides of polylinker in the 300 nucleotide antisense transcript can dramatically modify its effectiveness. The most effective antisense RNA construction was used in flow cytometry studies to investigate the role of ybr1012p. The results show that during the antisense RNA block some 80% of the cells are arrested with their DNA unreplicated, suggesting that Ybr1012p is needed for progression through G1 or early S phase.
YBR1012（YBR136w）是在酿酒酵母二号染色体右臂部分的系统测序过程中鉴定出的一个必需基因。我们之前基于反义RNA构建了YBR1012的条件等位基因，方法是将该基因的一小段片段插入可诱导的UASGAL10 - CYC1启动子下游。此后又构建了其他几种反义RNA结构并测试了它们的活性。该系统的反应似乎非常敏感，因为在300个核苷酸的反义转录本中多克隆位点13个核苷酸的有无会显著改变其有效性。最有效的反义RNA结构被用于流式细胞术研究，以探究ybr1012p的作用。结果表明，在反义RNA阻断期间，约80%的细胞因DNA未复制而停滞，这表明Ybr1012p是细胞通过G1期或早期S期所必需的。
