Gill Ryan T, Katsoulakis Eva, Schmitt William, Taroncher-Oldenburg Gaspar, Misra Jatin, Stephanopoulos Gregory
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
J Bacteriol. 2002 Jul;184(13):3671-81. doi: 10.1128/JB.184.13.3671-3681.2002.
We report the results of whole-genome transcriptional profiling of the light-to-dark transition with the model photosynthetic prokaryote Synechocystis sp. strain PCC 6803 (Synechocystis). Experiments were conducted by growing Synechocystis cultures to mid-exponential phase and then exposing them to two cycles of light/dark conditions, during which RNA samples were obtained. These samples were probed with a full-genome DNA microarray (3,169 genes, 20 samples) as well as a partial-genome microarray (88 genes, 29 samples). We concluded that (i) 30-min sampling intervals accurately captured transcriptional dynamics throughout the light/dark transition, (ii) 25% of the Synechocystis genes (783 genes) responded positively to the presence of light, and (iii) the response dynamics varied greatly for individual genes, with a delay of up to 120 to 150 min for some genes. Four classes of genes were identified on the basis of their dynamic gene expression profiles: class I (108 genes, 30-min response time), class II (279 genes, 60 to 90 min), class III (258 genes, 120 to 150 min), and class IV (138 genes, 180 min). The dynamics of several transcripts from genes involved in photosynthesis and primary energy generation are discussed. Finally, we applied Fisher discriminant analysis to better visualize the progression of the overall transcriptional program throughout the light/dark transition and to determine those genes most indicative of the lighting conditions during growth.
我们报告了以光合原核生物集胞藻PCC 6803株(集胞藻)为模型,对从光照到黑暗转变过程进行全基因组转录谱分析的结果。实验通过将集胞藻培养物培养至指数中期,然后使其经历两个光/暗循环来进行,在此期间获取RNA样本。这些样本用全基因组DNA微阵列(3169个基因,20个样本)以及部分基因组微阵列(88个基因,29个样本)进行检测。我们得出以下结论:(i)30分钟的采样间隔准确地捕捉了整个光/暗转变过程中的转录动态;(ii)25%的集胞藻基因(783个基因)对光照的存在有正向反应;(iii)各个基因的反应动态差异很大,一些基因的延迟长达120至150分钟。根据其动态基因表达谱鉴定出四类基因:I类(108个基因,反应时间30分钟)、II类(279个基因,60至90分钟)、III类(258个基因,120至150分钟)和IV类(138个基因,180分钟)。讨论了参与光合作用和初级能量产生的几个基因转录本的动态。最后,我们应用Fisher判别分析来更好地可视化整个光/暗转变过程中总体转录程序的进展,并确定那些最能指示生长期间光照条件的基因。