Watts F, Castle C, Beggs J
EMBO J. 1983;2(11):2085-91. doi: 10.1002/j.1460-2075.1983.tb01704.x.
We have investigated the ability of transcripts of the Drosophila melanogaster alcohol dehydrogenase gene to be spliced in Saccharomyces cerevisiae. The alcohol dehydrogenase gene was cloned in S. cerevisiae on a 2 micron DNA-based vector and a hybrid yeast actin-Drosophila alcohol dehydrogenase gene was constructed to demonstrate that transcripts encoded on a 2 micron plasmid could be accurately and efficiently spliced. Transcription of the Drosophila gene occurred in yeast with and without a yeast promoter. The transcripts were polyadenylated and terminated approximately 600 nucleotides distal to the polyadenylation site used in Drosophila. In yeast no splicing of the two introns within the alcohol dehydrogenase coding sequence was detected. However, the leader sequence was apparently spliced using the same 3' splice site as is used in adult flies, but a different 5' splice site. This result may be partly explained by the existence in the Drosophila gene of a sequence which is believed to be required for splicing in S. cerevisiae.
我们研究了果蝇乙醇脱氢酶基因的转录本在酿酒酵母中进行剪接的能力。乙醇脱氢酶基因被克隆到酿酒酵母中基于2μm DNA的载体上,并构建了一个酵母肌动蛋白-果蝇乙醇脱氢酶基因的杂种基因,以证明在2μm质粒上编码的转录本能够被准确且高效地剪接。果蝇基因的转录在有或没有酵母启动子的酵母中都能发生。转录本进行了多聚腺苷酸化,并且在果蝇中使用的多聚腺苷酸化位点下游约600个核苷酸处终止。在酵母中未检测到乙醇脱氢酶编码序列内两个内含子的剪接。然而,前导序列显然使用了与成年果蝇相同的3'剪接位点进行剪接,但5'剪接位点不同。这一结果可能部分归因于果蝇基因中存在一种被认为是酿酒酵母剪接所必需的序列。