笋瓜韧皮部蛋白基因启动子的克隆及其功能初探  

Cloning and Function of the Phloem Protein Gene Promoter from Cucurbita maxima

作　　者：蒋浩[1] 秦红敏[1] 虞红梅[1] 田颖川[1] 

机构地区：[1]中科院微生物研究所,北京100080

出　　处：《农业生物技术学报》1999年第1期63-68,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金;国际科学与文化中心(ICS)世界实验室的部分资助

摘　　要：笋瓜韧皮部蛋白（phloemprotein）PP2是一种独特的结合几丁质的凝集素，具有韧皮部组织特异性。为了研究该基因启动子在转基因植物中的表达特性，探索其在植物基因工程中潜在的应用价值，根据发表的序列，用PCR方法从笋瓜基因组中扩增得到了PP2基因启动子片段。经序列分析证明无误后，与GUS基因融合构建了植物表达载体。通过农杆菌介导转化烟草，经PCR分析初步表明嵌合基因已经整合到烟草基因组中。GUS组织化学检测显示转基因烟草的茎、叶柄和根韧皮部以及叶脉都为GUS染色阳性，这是首次用转基因植物证明笋瓜PP2基因启动子可驱动外源基回在异源植物韧皮部及分生组织中特异性表达。Phloem protein 2 (PP2) from Cucubita maxima is an unique chitin-binding lectin which shows phloemspecificity. In order to study the expression pattern in transgenic plants of its promotef, and tO explore its potential application in genetic engineering of plains, the promoter fragment of PP2 gene was amplified by PCR method from Cucurbita maxima genome and cloned. Intermediate vector was constructed by inserting the promoter to the upstream of GUS gene in pBI121,where the original CaMV 35S promoter was replaced by the PP2 promoter Regenerated tobacco plants were obtained by transformation mediated by Agrobacterium tumcfaciens. The result of PCR analysis indicated that the chimeric gene has probably been imegrated into the genome of regenerated plants. Histochemical localization of GUS activity indicated that, in transgenic tobacco plan, the C maxlina PP2 promoter could confer a phloem-and meristem-specific expression pattern to GUS gene.

关 键 词：笋瓜 韧皮部 特异性启动子 GUS嵌合基因 

分 类 号：S642.903.2[农业科学—蔬菜学]