Abstract：1,5 ribulose bisphosphate carboxylase/oxygenase (Rubisco) is the key enzyme of photosynthetic carbon assimilation in green plants, and its small structural subunits (Rubisco small subuni, RbcS) affect the function of Rubisco holoenzyme. In this study, the maize (Zea mays) inbred line B73 was material, DNA and RNA were firstly extracted from the leaves, the promoter sequence of ZmRbcS1 gene (GenBank No. 542212) was amplified and its structure was analyzed. Meanwhile, the ORF of the gene was amplified and the vector was constructed. Based on the analysis of the promoter by PlantCARE database, it was found that in addition to a large number of conservative light regulatory elements (such as G-box, GATA-motif, I-box, RbcS-CMA7c, etc.), the promoter sequence also contained a variety of conservative cis-acting elements (such as drought-induced response element (MBS) and gibberellin-responsive element (gibberellin-responsive element) in response to drought, hormones and other stress factors. P-box) and jasmonic acid response element (CGTCA-motif, TGACG-motif). According to the structure characteristics of ZmRbcS1 promoter and the distribution of regulating elements, three upstream primers and one downstream primer were designed for the amplification of promoter fragments of different lengths, with the lengths of 2 114, 1 021 and 566 bp, respectively. Subsequently, the 5' end of the promoter was deleted and the deletion expression vector of the reporter gene gap-glucuronidase (GUS) was constructed by fusing three segments of different lengths, and the promoter function of the instantaneous expression system of tobacco (Nicotiana benthamiana) leaves mediated by Agrobacterium tumefaciens was analyzed. Histochemical staining showed that the promoter activity increased first and then decreased with the reduction of promoter fragment. This study verified that the protein product encoded by ZmRbcS1 gene was located in the chloroplast, and the analysis and identification of its promoter clock-acting element provide the basis for further study on the function of ZmRbcS1 gene.
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