三角褐指藻甘油兼养条件下甘油激酶的功能研究  

Study on the function of glycerol kinase in Phaeodactylum tricornutumduring mixotrophic

作　　者：陈波[1] 黄龙[1] 郭婷[1] 卿人韦[1] 

机构地区：[1]四川大学生命科学学院,成都610064

出　　处：《四川大学学报（自然科学版）》2015年第1期144-148,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家863计划(2007AA09Z427);国家自然科学基金(40976092);中央高校项目(2010SCU22006)

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2079bp,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.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.

关 键 词：三角褐指藻:甘油激酶 兼养 RNAI 生物能源 

分 类 号：Q786[生物学—分子生物学]