Stockhaus J, Schell J, Willmitzer L
Institut für Genbiologische Forschung Berlin GmbH, FRG.
EMBO J. 1989 Sep;8(9):2445-51. doi: 10.1002/j.1460-2075.1989.tb08379.x.
A detailed analysis of the expression of a chimeric gene, consisting of the upstream region of the nuclear photosynthetic gene ST-LS1, encoding a component of the water-oxidizing complex of photosystem II, fused to the coding sequence of beta-glucuronidase (GUS) as a reporter, is described. The expression of this chimeric gene at the cellular level was detected by histochemical methods and shows that the expression of this gene is correlated with the presence of chloroplasts. Interestingly, the GUS activity was not only detected in typical photosynthetic tissues, e.g. leaves and stems, but also in green roots containing chloroplasts. In contrast no activity was detected in neighbouring white root tissue which was devoid of chloroplasts. One can therefore separate the relative importance of the (morphological) differentiation steps responsible for the formation of tissues normally involved in photosynthesis, from the importance of the developmental stage (characterized by the presence of chloroplasts), for the expression of this nuclear photosynthetic gene. Our data strongly suggest that the developmental stage of the plastids is the primary determinant for the activity of this nuclear photosynthetic gene, although they do not yet allow the exclusion of the reverse type of control, i.e. control of the differentiation of the plastid by the expression of certain nuclear genes. A chimeric gene, consisting of the promoter of the 35S cauliflower mosaic virus (CaMV) gene and the GUS coding sequence, was used as a control throughout the experiments, confirming that the observed differential ST-LS1-GUS gene expression reflects the particular transcriptional regulation impacted on this gene by its cis-acting regulatory sequences.
本文描述了对一个嵌合基因表达的详细分析,该嵌合基因由光合核基因ST-LS1的上游区域与β-葡萄糖醛酸酶(GUS)的编码序列融合而成,其中ST-LS1编码光系统II水氧化复合物的一个组分,GUS作为报告基因。通过组织化学方法检测了该嵌合基因在细胞水平的表达,结果表明该基因的表达与叶绿体的存在相关。有趣的是,不仅在典型的光合组织如叶片和茎中检测到了GUS活性,在含有叶绿体的绿色根中也检测到了活性。相反,在不含叶绿体的相邻白色根组织中未检测到活性。因此,可以将负责形成通常参与光合作用的组织的(形态学)分化步骤的相对重要性,与发育阶段(以叶绿体的存在为特征)对该光合核基因表达的重要性区分开来。我们的数据强烈表明,质体的发育阶段是该光合核基因活性的主要决定因素,尽管目前还不能排除相反类型的控制,即某些核基因的表达对质体分化的控制。在整个实验中,使用了一个由35S花椰菜花叶病毒(CaMV)基因的启动子和GUS编码序列组成的嵌合基因作为对照,证实了观察到的ST-LS1-GUS基因的差异表达反映了其顺式作用调控序列对该基因的特定转录调控。