磷转运蛋白基因表达载体的构建及对烟草的转化.docVIP

 磷转运蛋白基因表达载体的构建及对烟草 的转化 张洁莉，韩阳** 5 10 （辽宁大学生命科学院，沈阳 110036） 摘要：磷是植物生长过程中重要的营养元素，调控植物的整个代谢过程,而土壤中能供给植 物吸收的有效磷甚少，满足不了植物的需求，成为阻碍植物生长发育的因素之一。磷元素在 植物体内的流动是一个复杂的过程，它的吸收和转运主要由磷转运蛋白负责。在缺磷的环境 中，植物会通过提高磷转运蛋白基因的表达量来促进植物根系及其共生菌根菌丝对磷的吸收 及转运。将克隆的番茄磷酸转运蛋白基因 LePT1 插入 pCAMBIA2300，构建成植物表达载体。 采用农杆菌介导法转化烟草，获得转基因植株。对转基因植株的 PCR 分析表明，外源基因 已整合进转基因植株中并得到表达。 关键词：磷转运蛋白基因；表达载体；遗传转化；烟草 中图分类号：Q94 15 Vector construction of phosphate transporter gene expression vector and transformation Tobacco Zhang Jieli, Han Yang (School of Life Sciences,Liaoning University, ShenYang 110036) 20 25 30 35 40 Abstract: Phosphorus is an important plant nutrient, and it regulates entire metabolic processes. However, the content of phosphate available to plants in soil is very low, and it often cannot meet the requirements of plants. Therefore, the available phosphate supply in soil and the ability of plants to absorb phosphate is one of determinants of plant growth and development. Phosphate mobilization into the plant is a complex process in which the absorption and translocation of this major nutrient are determined largely by the phosphate transporters. Plants regulate the expression of Pi transporter genes to adapt low available P environment by enhancing their ability to acquire and transfer Pi. A plant expression vector was constructed by inserting the phosphate transporter gene, LePT1, which was cloned from the tomato genome, into pCAMBIA2300. An agrobacterium-mediated system was used to transform tobacco and acquire transgenic plants. Analyses of the transgenic plants by PCR indicated that the exogenous gene was integrated into and expressed by transgenic plants. Keywords: phosphate transporter genes; expression vector; transformation; tobacco 磷是植物生长过程中重要的营养元素，调控植物的整个代谢过程。而土壤中能供给植物 吸收的有效磷甚少，满足不了植物的需求。土壤有效磷的供应状况和植物对磷素营养的吸收 能力便成为植物生长发育的主要决定因素之一[1]。 磷的吸收主要通过细胞膜上的磷转运系统进行，目前已经发现的磷转运子 Pht1 家族， 其成员主要负责植物根系对磷的吸收及转运，表达受磷的调控[2]。LePT1 基因是来源于番茄， 编码高亲和力磷转运蛋白，属于 Phtl 家族的成员。本实验以 LePT1 基因为目的基因，构建 植物表达载体，将 LePT1 转入模式