Microalgal lipid triacylglycerol (TAG) is a promising source for sustainable production of biofuels and edible oils. TAG biosynthesis in microalgae can be induced by nitrogen starvation (−N); however, regulation of the genes involved in this process is poorly known. To explore the regulation of gene encoding diacylglycerol acyltransferase 2 of oleaginous microalga Neochloris oleoabundans (NeoDGAT2) responsible for TAG biosynthesis, regulatory sequence of NeoDGAT2 gene (RDG) was cloned, and its functional regions were mapped by deletion analysis using the modified cyan fluorescent protein gene (CFP) as a reporter. The efficiency of CFP gene mTurquoise2 (Tq) without any intron, Tq1 and Tq2 with one and two copies of Chlamydomonas reinhardtii rbcS2 intron 1, respectively, was evaluated; Tq2 exhibited the highest CFP fluorescence activity in N. oleoabundans was therefore used as reporter for RDG deletion analysis. Deletion analysis of RDG revealed that the −N inducible region contained the predicted binding site of MYB transcription factor (MYB-bs). Specific binding between MYB-bs of RDG and the DNA-binding domain of MYB-related transcription factor ROC40 from C. reinhardtii was observed using electrophoretic mobility shift assay. Therefore, the corresponding MYB transcription factor in N. oleoabundans is probably the transcription factor regulating NeoDGAT2. The interaction between MYB transcription factor and the MYB-bs may play a role in regulating −N induced expression of NeoDGAT2, affecting TAG accumulation. MYB transcription factors can be the potential targets for engineering to increase TAG content. Increasing TAG content is essential for products derived from microalgal TAG to achieve economic viability.

微藻脂质三酰基甘油（TAG）是可持续生产生物燃料和食用油的有希望的来源。氮饥饿（-N）可以诱导微藻中的TAG生物合成；然而，对该过程涉及的基因的调控知之甚少。为了探索负责TAG生物合成的含油微藻Neochloris oleoabundans（NeoDGAT2）的编码二酰基甘油酰基转移酶2的基因的调控，克隆了NeoDGAT2基因（RDG）的调控序列，并使用修饰的蓝绿色荧光蛋白基因通过缺失分析来定位其功能区（CFP）作为记者。的效率CFP基因mTurquoise2（Tq的），不含任何内含子，TQ1和TQ2具有一个和两个拷贝莱茵衣藻rbcS2的内含子1，分别进行评价; TQ2表现出最高的CFP荧光活性N. oleoabundans因此用作记者为RDG缺失分析。RDG的缺失分析表明，-N诱导区含有MYB转录因子（MYB-bs）的预测结合位点。RDG的MYB-b与来自莱茵衣藻的MYB相关转录因子ROC40的DNA结合结构域之间的特异性结合使用电泳迁移率变动分析法观察到。因此，油橄榄猪笼草中相应的MYB转录因子可能是调节NeoDGAT2的转录因子。MYB转录因子与MYB-bs之间的相互作用可能在调节-N诱导的NeoDGAT2表达中发挥作用，影响TAG的积累。MYB转录因子可能是工程设计以增加TAG含量的潜在目标。TAG含量的增加对于衍生自微藻TAG的产品实现经济可行性至关重要。