ASPEX Division, Research Center, Asahi Glass Co, Ltd, Yokohama, Kanagawa 221-8755, Japan.
Nucleic Acids Res. 2013 May 1;41(10):5382-99. doi: 10.1093/nar/gkt233. Epub 2013 Apr 5.
The Schizosaccharomyces pombe genome is one of the smallest among the free-living eukaryotes. We further reduced the S. pombe gene number by large-scale gene deletion to identify a minimal gene set required for growth under laboratory conditions. The genome-reduced strain has four deletion regions: 168.4 kb in the left arm of chromosome I, 155.4 kb in the right arm of chromosome I, 211.7 kb in the left arm of chromosome II and 121.6 kb in the right arm of chromosome II. The deletions corresponded to a loss of 223 genes of the original ~5100. The quadruple-deletion strain, with a total deletion size of 657.3 kb, showed a decreased ability to uptake glucose and some amino acids in comparison with the parental strain. The strain also showed increased gene expression of the mating pheromone M-factor precursor and the nicotinamide adenine dinucleotide phosphate -specific glutamate dehydrogenase. There was also a 2.7-fold increase in the concentration of cellular adenosine triphosphate, and levels of the heterologous proteins, enhanced green fluorescent protein and secreted human growth hormone were increased by 1.7- and 1.8-fold, respectively. The transcriptome data from this study have been submitted to the Gene Expression Omnibus (GEO: http://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE38620 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=vjkxjewuywgcovc&acc=GSE38620).
裂殖酵母的基因组是自由生活真核生物中最小的基因组之一。我们通过大规模基因缺失进一步减少了裂殖酵母的基因数量,以鉴定在实验室条件下生长所需的最小基因集。基因组减少的菌株有四个缺失区域:I 号染色体左臂的 168.4kb、I 号染色体右臂的 155.4kb、II 号染色体左臂的 211.7kb 和 II 号染色体右臂的 121.6kb。缺失对应于原始约 5100 个基因中的 223 个基因的丢失。与亲本菌株相比,四重缺失菌株(总缺失大小为 657.3kb)摄取葡萄糖和一些氨基酸的能力下降。该菌株还表现出交配信息素 M 因子前体和烟酰胺腺嘌呤二核苷酸磷酸特异性谷氨酸脱氢酶的基因表达增加。细胞三磷酸腺苷的浓度也增加了 2.7 倍,异源蛋白增强型绿色荧光蛋白和分泌型人生长激素的浓度分别增加了 1.7 倍和 1.8 倍。本研究的转录组数据已提交给基因表达综合数据库(GEO:http://www.ncbi.nlm.nih.gov/geo/),登录号为 GSE38620(http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=vjkxjewuywgcovc&acc=GSE38620)。