Yu HaoQiang, Khalid Muhammad Hayder Bin, Lu FengZhong, Sun FuAi, Qu JingTao, Liu BingLiang, Li WanChen, Fu FengLing
Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region, Ministry of Agriculture; Maize Research Institute, Sichuan Agricultural University, Chengdu, 611130 People's Republic of China.
Physiol Mol Biol Plants. 2019 Jan;25(1):277-287. doi: 10.1007/s12298-018-0546-z. Epub 2018 Jun 4.
To avoid the unregulated overexpression of the exogenous genes, specific or inducible expression is necessary for some exogenous genes in transgenic plants. But little is known about organ- or tissue-specific promoters in maize. In the present study, the expression of a maize pentatricopeptide repeat (PPR) protein encoding gene, , was analyzed by RNA-sequencing data and confirmed by quantitative real time PCR. The results showed that the gene specifically expressed in vegetative organs. Consequently, a 1830 bp sequence upstream of the start codon of the promoter for gene was isolated from maize genome ( ). To validate whether the promoter possesses the vegetative organ-specificity, the full-length and three 5'-end deletion fragments of of different length (1387, 437, and 146 bp) were fused with glucuronidase (GUS) gene to generate constructs and transformed into tobacco. The transient expression and fluorometric GUS assay in transgenic tobacco showed that all promoter could drive the expression of the gene, the - 437 to - 146 bp region possessed some crucial elements for root-specific expression, and the shortest and optimal sequence to maintain transcription activity was 146 and 437 bp in length, respectively. These results indicate that the promoter of the gene is a vegetative organ-specific promoter and will be useful in transgenic modification of commercial crops for moderate specific expression after further evaluation in monocotyledons.
为避免外源基因的无节制过表达,转基因植物中某些外源基因需要特异性或诱导性表达。但关于玉米中器官或组织特异性启动子的了解甚少。在本研究中,通过RNA测序数据分析了一个玉米五肽重复(PPR)蛋白编码基因的表达,并通过定量实时PCR进行了验证。结果表明,该基因在营养器官中特异性表达。因此,从玉米基因组中分离出该基因启动子起始密码子上游1830 bp的序列( )。为验证该启动子是否具有营养器官特异性,将不同长度(1387、437和146 bp)的该基因全长和三个5'端缺失片段与葡萄糖醛酸酶(GUS)基因融合,构建 载体并转化到烟草中。转基因烟草中的瞬时表达和荧光GUS分析表明,所有启动子均可驱动该基因的表达,-437至-146 bp区域具有一些根特异性表达的关键元件,维持转录活性的最短和最佳序列长度分别为146和437 bp。这些结果表明,该基因的启动子是一种营养器官特异性启动子,在单子叶植物中进一步评估后,将有助于商业作物的转基因改良以实现适度的特异性表达。