Xu Bi-Yu, Liu Ge, Jin Zhi-Qiang
State Key Laboratory of Tropical Crops Biotechnology, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China.
Sheng Wu Gong Cheng Xue Bao. 2006 Nov;22(6):945-9.
Banana (Musa spp) is one of the most important fruit crops in the world. Banana fruit is an ideal organ for producing foreign pharmaceutical proteins and chemicals by genetic engineering. A perfect promoter driving foreign gene to express strongly and specifically in banana fruit is necessary for that. In order to isolate a banana fruit-specific expressed promoter, a fragment of 702 nt nucleotide sequence upstream 5' of banana lectin (BanLec) gene, which was demonstrated to express specifically in banana fruit previously, was isolated by using chromosomal walking in this study. Bioinformatical analysis of this sequence shows that the sequence contains some typical elements of a promoter. To identify the fruit-specific expression of this promoter, a construct was derived from pBI121, which originally CaMV 35S promoter was replaced by the 702 nt nucleotide sequence, and named as pBIL2. Transformations of pBIL2 to roots, leaves and fruit pieces of banana were carried out by using particle bombardment. The transient expression of gus showed that the gus expressed specifically in banana fruit with a little higher level compared with CaMV 35S. It is the first report that BanLec promoter is a potential fruit-specific expressed promoter which can further be used in transgenes into banana.
香蕉(芭蕉属)是世界上最重要的水果作物之一。香蕉果实是通过基因工程生产外源药用蛋白和化学物质的理想器官。为此,需要一个能驱动外源基因在香蕉果实中强烈且特异性表达的完美启动子。为了分离出香蕉果实特异性表达的启动子,本研究利用染色体步移技术,分离出了香蕉凝集素(BanLec)基因5'端上游702 nt的核苷酸序列片段,该片段先前已被证明在香蕉果实中特异性表达。对该序列的生物信息学分析表明,该序列包含启动子的一些典型元件。为了鉴定该启动子的果实特异性表达,构建了一个源自pBI121的载体,将其原有的花椰菜花叶病毒35S启动子替换为702 nt的核苷酸序列,并命名为pBIL2。通过粒子轰击将pBIL2转化到香蕉的根、叶和果实切片中。gus的瞬时表达表明,gus在香蕉果实中特异性表达,且表达水平略高于花椰菜花叶病毒35S启动子。这是首次报道BanLec启动子是一种潜在的果实特异性表达启动子,可进一步用于香蕉的转基因研究。