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 tricornutum during mixotrophic
作者：陈波(四川大学生命科学学院)；黄龙(四川大学生命科学学院)；郭婷(四川大学生命科学学院)；卿人韦(四川大学生命科学学院)
Author：CHEN Bo(College of Life Sciences, Sichuan University)；HUANG Long(College of Life Sciences, Sichuan University)；GUO Ting(College of Life Sciences, Sichuan University)；QING Ren-Wei(College of Life Sciences, Sichuan University)
收稿日期：2013-05-14 年卷（期）页码：2015,52(1):144-148
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：三角褐指藻: 甘油激酶; 兼养; RNAi; 生物能源
Key words：Phaeodactylum tricornutum, glycerol kinase, mixotrophic, RNAi, Biodiesel
基金项目：国家863计划(2007AA09Z427)；国家自然科学基金(40976092)；中央高校项目(2010SCU22006)
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 constructed based on the ORF of PtrGK and transformed to the E. coli BL21 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 transformants 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 glycerol kinase in Phaeodactylum tricornutum during mixotrophic cultivate and have a reference value in improving the diatom cell division rate during glycerol mixotrophic.
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