三角褐指藻(Phaeodactylum tricornutum)通用转化载体的构建

摘要

Phaeodactylum tricornutum is an ideal material for biodiesel research. In this paper, several endogenous elements, i.e. promoters and terminators, fromfcp (fucoxanthin, chlorophyll a/c-binding protein) gene clusters were cloned, and a transformation vector pfcpA-MCS/fcpB-bar including two expression cassettes which were fcpB promoter-bar gene-fcpA terminatiom and fcpA promoterm-MCS (multiple cloning site)-fcpA terminatiom was constructed and characterized by using bar gene as selection marker. It carried a large MCS easy for insertion of one or more function genes. This vector will be used for lipid metabolic engineering for Phaeodactylum tricorn-utum.%三角褐指藻(Phaeodactylum tricornutum)是开展微藻生物柴油研究的理想材料.克隆了内源fcp基因簇的多个调控序列(启动子、终止子),构建了包括fcpB启动子-bar基因-fcpA终止子、以及fcpA启动子-多克隆位点(MCS)-fcpA终止子两个表达盒的通用转化载体pfcpA-MCS/fcpB-Bar,其特征是以bar基因作为选择标记,MCS区方便插入一至多个目的基因.新载体可用于三角褐指藻的重组蛋白表达、或油脂代谢相关基因的功能验证和代谢调控研究.