Singh A K, Sherman L A
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
J Bacteriol. 2000 Jun;182(12):3536-43. doi: 10.1128/JB.182.12.3536-3543.2000.
We describe the use of a method called differential expression using customized amplification library (DECAL) to study the global changes in gene expression in iron-deficient versus iron-reconstituting cells of Synechocystis sp. strain PCC 6803. We identified a number of genes, such as isiA, idiA, psbA, cpcG, and slr0374, whose expression either increased or decreased in response to iron availability. Further analysis led to the identification of additional genes related to those identified by DECAL (e.g., psbC, psbO, psaA, apcABC, cpcBAC1C2D, and nblA) that were differentially regulated by iron availability. Expression of cpcG, psbC, psbO, psaA, apcABC, and cpcBAC1C2D increased, whereas that of isiA, idiA, nblA, psbA, and slr0374 decreased, in iron-reconstituting cells. S1 nuclease protection studies showed that increased transcript levels of psbA in iron-deficient cells was due to the increased expression of both psbA2 and psbA3 genes, although the steady-state level of psbA2 remained higher than that of psbA3.
我们描述了一种名为使用定制扩增文库进行差异表达(DECAL)的方法的应用,以研究集胞藻PCC 6803菌株缺铁细胞与铁重构细胞中基因表达的整体变化。我们鉴定了许多基因,如isiA、idiA、psbA、cpcG和slr0374,它们的表达随铁供应情况而增加或减少。进一步分析导致鉴定出与DECAL鉴定的基因相关的其他基因(如psbC、psbO、psaA、apcABC、cpcBAC1C2D和nblA),这些基因受铁供应情况的差异调节。在铁重构细胞中,cpcG、psbC、psbO、psaA、apcABC和cpcBAC1C2D的表达增加,而isiA、idiA、nblA、psbA和slr0374的表达减少。S1核酸酶保护研究表明,缺铁细胞中psbA转录水平的增加是由于psbA2和psbA3基因表达均增加,尽管psbA2的稳态水平仍高于psbA3。