The Preparation And Identification Of RbcS3A-gus And RbcS3C-gus Transgenic Tobacco And Tomato Plants

The rbcS was encoded by a gene family in plants. Light was one of important factors to regulate its gene expression. Meanwhile, rbcS gene also expressed in a light-independent manner. However, the mechanism for the later is not clear yet. Our preliminary experiments suggested that extracellular calmodulin might be involved in rbcS gene expression in the dark.In order to provide direct evidence that extracellular calmodulin activated rbcS transcription in a light-independent manner, we intended to get the transgenic plants carrying light-dependent and -independent rbcS promoter fused to a reporter gene.To do so, we first selected two tomato rbcS genes, rbcS3A and rbcS 3C, to represent light-independent and -dependent expression member of this gene family, respectively, and amplified the promoter region of those two genes from tomato genome. Then, we transformed those two genes into tomato and tobacco plants via Agrobacterium tumefaciens. After verified by antibiotic resistance, reporter gene examination, southern blot detection, and genetic segregation analysis, we obtained 3 and 7 transgenic tobacco plants with one copy of rbcS 3A-gus or rbcS 3C-gus gene, respectively.Further, we established two suspension-cultured cell lines using above mentioned two kinds of transgenic tobacco plants.