Guerrero F D, Crossland L, Smutzer G S, Hamilton D A, Mascarenhas J P
Ciba-Geigy Biotechnology Center, Research Triangle Park, NC 27709.
Mol Gen Genet. 1990 Nov;224(2):161-8. doi: 10.1007/BF00271548.
A set of 5' promoter deletions from Zmg13, a genomic clone of a pollen-specific gene of maize, has been transcriptionally fused to a beta-glucuronidase (GUS) reporter gene in the binary vector pBI101. Tobacco leaf disks were transformed and mature plants analyzed for GUS activity directed by the Zmg13 promoter constructs. Transgenic plants containing the 375 bp Zmg13 sequence from -314 to +61 relative to the transcription start site transcribed GUS RNA and expressed active GUS enzyme in mature pollen but not in leaves. Plants transformed with a 35S CaMV promoter-GUS transcriptional fusion expressed GUS RNA in leaves but not in pollen. Neither GUS RNA or active enzyme could be detected in pollen or leaves from plants containing a 124 bp Zmg13-GUS transcriptional fusion missing the putative Zmg13 TATA box. No GUS RNA or enzyme expression was not detected in non-transformed tobacco. RNA and GUS histochemical analysis of the T1 generation confirmed that the temporal expression pattern of Zmg13-GUS transcription in tobacco followed that of the native gene in maize and that the Zmg13 promoter sequences from the maize gene are able correctly to direct genetically stable, tissue-specific gene expression in transgenic tobacco plants.
从玉米花粉特异性基因的基因组克隆Zmg13中获得了一组5'端启动子缺失片段，这些片段已在二元载体pBI101中与β-葡萄糖醛酸酶（GUS）报告基因进行转录融合。对烟草叶盘进行转化，并对成熟植株分析由Zmg13启动子构建体指导的GUS活性。含有相对于转录起始位点从-314到+61的375 bp Zmg13序列的转基因植物在成熟花粉中转录GUS RNA并表达活性GUS酶，但在叶片中不表达。用35S CaMV启动子-GUS转录融合体转化的植物在叶片中表达GUS RNA，但在花粉中不表达。在含有缺失假定的Zmg13 TATA框的124 bp Zmg13-GUS转录融合体的植物的花粉或叶片中，均未检测到GUS RNA或活性酶。在未转化的烟草中未检测到GUS RNA或酶表达。对T1代的RNA和GUS组织化学分析证实，烟草中Zmg13-GUS转录的时间表达模式与玉米中天然基因的表达模式一致，并且来自玉米基因的Zmg13启动子序列能够正确地指导转基因烟草植株中遗传稳定的组织特异性基因表达。
