Sazuka T, Ohara O
Laboratory of DNA Technology, Kazusa DNA Research Institute, Chiba, Japan.
Electrophoresis. 1997 Aug;18(8):1252-8. doi: 10.1002/elps.1150180806.
Following the complete sequencing of the genome of the univellular cyanobacterium, Synechocystis sp. strain PCC6803 within our institute, a protein-gene linkage map of this photosynthetic microorganism was successfully constructed for 130 high abundance proteins present on two-dimensional gels. An additional six proteins were analyzed, but were probably encoded extrachromosomally. In order to demonstrate the usefulness of this protein-gene linkage map, we analyzed the changes that occur in cellular proteins after illumination of PCC6803 cells. The results indicate that this protein-gene linkage map greatly simplifies the identification process of such modulated genes. After illumination, at least three distinctive spots with reduced intensity were detected on two-dimensional gels and the corresponding genes of two of these were successfully identified as chaperonin 2 and a Tortula ruralis rehydrin-related gene. Thus, the combination of the protein-gene linkage map and two-dimensional gel electrophoresis should permit a comprehensive analyses of the proteins encoded by the genome (i.e., "proteome") of this photosynthetic autotroph. This post-genome project represents a productive way of exploiting the information obtained from the sequencing of the cyanobacterium genome.
在我们研究所完成单细胞蓝藻聚球藻属PCC6803菌株的基因组全序列测定之后,针对二维凝胶上存在的130种高丰度蛋白质成功构建了这种光合微生物的蛋白质-基因连锁图谱。另外分析了6种蛋白质,但它们可能是由染色体外编码的。为了证明这种蛋白质-基因连锁图谱的实用性,我们分析了PCC6803细胞光照后细胞内蛋白质发生的变化。结果表明,这种蛋白质-基因连锁图谱极大地简化了此类调控基因的鉴定过程。光照后,在二维凝胶上检测到至少三个强度降低的明显斑点,其中两个斑点的相应基因已成功鉴定为伴侣蛋白2和一个与卷叶藓脱水素相关的基因。因此,蛋白质-基因连锁图谱与二维凝胶电泳的结合应能对这种光合自养生物基因组(即“蛋白质组”)所编码的蛋白质进行全面分析。这个后基因组计划是利用从蓝藻基因组测序中获得信息的一种有效方式。