Onizuka Takuo, Endo Sumiyo, Akiyama Hideo, Kanai Shozo, Hirano Masahiko, Yokota Akiho, Tanaka Satoshi, Miyasaka Hitoshi
Biological Science Laboratories, Toray Research Center, Inc., 1111 Tebiro, Kamakura, Kanagawa, 248-8555 Japan.
Plant Cell Physiol. 2004 Oct;45(10):1390-5. doi: 10.1093/pcp/pch160.
The operon encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the cyanobacterium Synechococcus sp. PCC7002 contains three rbc genes, rbcL, rbcX and rbcS, in this order. Introduction of translational frameshift into the rbcX gene resulted in a significant decrease in the production of large (RbcL) and small (RbcS) subunits of the Rubisco protein in Synechococcus sp. PCC7002 and in Escherichia coli. To investigate the function of the rbcX gene product (RbcX), we constructed the expression plasmid for the rbcX gene and examined the effects of RbcX on the recombinant Rubisco production in Escherichia coli. The coexpression experiments revealed that RbcX had marked effects on the production of large and small subunits of Rubisco without any significant influence on the mRNA level of rbc genes and/or the post-translational assembly of the Rubisco protein. The present rbcX coexpression system provides a novel and useful method for investigating the Rubisco maturation pathway.
蓝藻聚球藻属(Synechococcus sp.)PCC7002中编码1,5-二磷酸核酮糖羧化酶/加氧酶(Rubisco)的操纵子依次包含三个rbc基因,即rbcL、rbcX和rbcS。在rbcX基因中引入翻译移码导致聚球藻属PCC7002和大肠杆菌中Rubisco蛋白的大亚基(RbcL)和小亚基(RbcS)产量显著下降。为了研究rbcX基因产物(RbcX)的功能,我们构建了rbcX基因的表达质粒,并检测了RbcX对大肠杆菌中重组Rubisco产量的影响。共表达实验表明,RbcX对Rubisco大亚基和小亚基的产量有显著影响,而对rbc基因的mRNA水平和/或Rubisco蛋白的翻译后组装没有任何显著影响。目前的rbcX共表达系统为研究Rubisco成熟途径提供了一种新颖且有用的方法。