中文摘要：

本研究利用RT-PCR结合 RACE 方法扩增出了三角褐指藻甘油激酶（PtrGK）基因全长 cDNA序列,其完整 ORF为2079 bp,编码692个氨基酸。基于上述序列构建了甘油激酶原核表达载体。构建反向互补RNA干扰载体并转化野生型三角褐指藻,得到含有甘油激酶反向互补干扰结构的转基因藻。研究表明：其甘油激酶表达水平、利用某油速率和细胞分裂速度都较野生型藻有了较大程度的下降。说明甘油激酶对三角褐指藻甘油兼养生长的重要作用。

英文摘要：

The full length of glycerol kinase （PtrGK）was isolated from Phaeodactylum tricornutum by using RT-PCR and RACE methods.Sequence analysis indicated that this gene has an open reading frame of 2079 bp,encoding 692 amino acids.Prokaryotic expression and invert-repeat RNAi vectors were con-structed based on the ORF of PtrGK and transformed to the E.coli BL2 1 and the diatom,respectively. Then obtain the glycerol kinase protein purified by Ni-NTA and RNAi transgenetic algae.Real-time PCR analysis revealed that there was a significantly lower expression level of the PtrGK in the transfor-mants than the wild type.And there was a lower rate of cell division and utilize the exogenous glycerol in genetically modified algae than the wild strain.The results revealed that the importance of the glycer-ol kinase in Phaeodactylum tricornutum during mixotrophic cultivate and have a reference value in im-proving the diatom cell division rate during glycerol mixotrophic.