盐藻的无菌培养及其抗生素标记的选择

Culture Condition and Selective Marker of Dunaliella salina

对影响盐藻生长的NaNO3、NaH2PO4、NaHCO3和VB1、VB12、VH等几种主要营养盐进行了优化,在单因素实验的基础上做了四因素三水平正交实验,实验结果得出优化配方为N/P值固定为f/2培养基,NaNO3和NaH2PO4添加量为f/2培养基的10倍、NaHCO30.4 g/L、VB1100μg/L、VB121.0μg/L,其他元素按f/2培养基添加.对盐藻的选择标记进行了研究,选择了氯霉素、G418、潮霉素3种抗生素进行实验,得出氯霉素适合作为盐藻基因工程的筛选抗生素,CAT基因为其阳性筛选标记基因,固体培养基筛选浓度为80μg/mL.为该藻的进一步高密度大规模培养和分子水平的研究提供了依据.

The effects of nutrient compositions such as nitrogen, phosphorus, carbon, vitamins B1 and B12 on the growth of axenic Dunaliella salina are studied. The concentrations of nutrient compositions are optimized by orthogonal experiment as follows ：the concentrations of NaNO3 and NaH2PO4 are taken 10 times as that of f/ 2 medium, N/P keeps the same concentration as f/2 medium, NaHCO3 0. 4 g/L, VB1 100 μg/L and VB12= 0. 5 μg/L. The same concentrations are chosen as that of f/2 medium for the others. Sensitivities of D. salina to chloromycetin（Cm） ,G418, and hygromycin are discussed. The results show that D. salina is sensitive to Cm, whose selective concentration in solid medium is 80μg/mL. It suggests that CAT gene could be a suitable selective marker for D. salina genetic engineering.