莱茵衣藻三基因共调控油脂积累研究

Study on the regulation of oil accumulation by three genes of Chlamydomonas reinhardtii

目的:通过对长链酰基辅酶A合成酶(LACS2)、柠檬酸合酶(CIS1)和转录因子(DOF)的共调控,构建莱茵衣藻(Chlamydomonas Reinhardtii)的高油脂积累三基因藻株.方法:将双基因沉默载体(LACS2-CIS1)转入DOF工程藻中得到三基因藻株,分析了其生长状况、淀粉、蛋白质及脂肪酸含量.结果:与莱茵衣藻cc849相比,三基因藻株生长状态相似,淀粉和蛋白无明显改变,胞内中性脂含量明显增加,且脂肪酸含量上升了31%.在热激启动作用下,三基因藻株淀粉和蛋白下降了58%和26%,胞内中性脂含量增加74%,脂肪酸含量上升了52%.结论:三基因共调控可促进莱茵衣藻胞内油脂积累.

Objective:Through the co-regulation of long-chain acyl-CoA synthase(LACS2),citrate synthase(CIS1)and transcription factor(DOF),a three-gene algal strain with high oil accumulation in Chlamydomonas reinhardtii was constructed.Methods:The double-gene silencing vector(LACS2-CIS1)was transferred into DOF engineering algae to obtain a three-gene algae strain,and its growth status,starch,protein and fatty acid content were analyzed.Results:Compared with Chlamydomonas reinhardtii cc849,the growth status of the three-gene algae strain was similar,the starch and protein did not change significantly,the intracellular neutral lipid content increased,and the fatty acid content increased by 31%.Under the effect of heat shock,the starch and protein of the three-gene algal strain decreased by 58%and 26%,the intracellular neutral lipid content increased by 74%,and the fatty acid content increased by 52%.Conclusion:Co-regulation of three genes can promote the accumulation of intracellular lipids in Chlamydomonas reinhardtii.