Kjaerulff Søren, Jensen Martin Roland
Pharmexa A/S, Kogle Allé 6, DK-2970, Hørsholm, Denmark.
Biochem Biophys Res Commun. 2005 Oct 28;336(3):974-82. doi: 10.1016/j.bbrc.2005.08.195.
In the fission yeast Schizosaccharomyces pombe, there are relatively few signal peptides available and most reports of their activity have not been comparative. Using sequence information from the S. pombe genome database we have identified three putative signal peptides, designated Cpy, Amy and Dpp, and compared their ability to support secretion of green fluorescent protein (GFP). In the comparison we also included the two well-described secretion signals derived from the precursors of, respectively, the Saccharomyces cerevisiae alpha-factor and the S. pombe P-factor. The capability of the tested signal peptides to direct secretion of GFP varied greatly. The alpha-factor signal did not confer secretion to GFP and all the produced GFP was trapped intracellular. In contrast, the Cpy signal peptide supported efficient secretion of GFP with yields approximating 10 mg/L. We also found that the use of an attenuated version of the S. cerevisiae URA3 marker substantially increases vector copy number and expression yield in fission yeast.
在裂殖酵母粟酒裂殖酵母中，可用的信号肽相对较少，并且关于它们活性的大多数报道都没有进行比较。利用粟酒裂殖酵母基因组数据库中的序列信息，我们鉴定出了三种假定的信号肽，分别命名为Cpy、Amy和Dpp，并比较了它们支持绿色荧光蛋白（GFP）分泌的能力。在比较中，我们还纳入了分别来源于酿酒酵母α-因子前体和粟酒裂殖酵母P-因子的两种已充分描述的分泌信号。所测试的信号肽指导GFP分泌的能力差异很大。α-因子信号不能使GFP分泌，所有产生的GFP都被困在细胞内。相反，Cpy信号肽支持GFP的有效分泌，产量约为10 mg/L。我们还发现，使用酿酒酵母URA3标记的弱化版本可显著增加裂殖酵母中的载体拷贝数和表达产量。
