Pasquali G, Ouwerkerk P B, Memelink J
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, The Netherlands.
Gene. 1994 Nov 18;149(2):373-4. doi: 10.1016/0378-1119(94)90179-1.
A convenient vector system was developed to evaluate transcriptional promoter activities in plants. Two primary vectors, optionally containing the cauliflower mosaic virus (CaMV) 35S -47 or -90 minimal promoters, offer multiple sites for cloning the sequence of interest upstream from the beta-glucuronidase gene (gusA). The promoter-gusA cassette can be transferred to a binary vector containing the selectable neomycin phosphotransferase II-encoding gene (nptII) next to the left border. In addition, the transferred DNA (T-DNA) contains the chloramphenicol acetyltransferase gene (cat) driven by the CaMV 35S promoter. Activity of cat can serve as a reference for gusA expression to correct for effect of chromosomal position or T-DNA copy number.
开发了一种便捷的载体系统来评估植物中的转录启动子活性。两种主要载体，可选择包含花椰菜花叶病毒（CaMV）35S -47或-90最小启动子，为在β-葡萄糖醛酸酶基因（gusA）上游克隆感兴趣的序列提供了多个位点。启动子-gusA盒可转移至一个二元载体，该二元载体在左边界旁含有编码新霉素磷酸转移酶II的基因（nptII）。此外，转移DNA（T-DNA）包含由CaMV 35S启动子驱动的氯霉素乙酰转移酶基因（cat）。cat的活性可作为gusA表达的参考，以校正染色体位置或T-DNA拷贝数的影响。
