转基因集胞藻PCC6803培养条件的优化

Optimization of Culture Conditions for Transgenic Synechocystis sp.PCC6803

通过研究培养基中硝酸盐、磷酸盐、碳酸盐等单因子对转牙鲆生长激素基因的集胞藻PCC6803生长的影响,再对以上3营养因子进行三水平的正交实验,得到培养基中硝酸盐、磷酸盐、碳酸氢盐的优化组合。还对培养温度、光照强度、pH值进行三因素三水平的正交实验,以确定最适的培养条件。实验结果表明,转基因集胞藻PCC6803在30℃,光照强度3 000lx,pH值8培养条件下,培养基中NaNO31.25 g/L,K2HPO40.04 g/L,Na2CO30.03 g/L(其它同BG-11培养基),藻细胞的生物量得到有效提高,特定生长速率达到20.64,这为转牙鲆生长激素基因集胞藻PCC6803进一步高密度放大培养提供了依据。

By examining the effects of nitrate,phosphate and carbonate contents in the medium on the growth of transgenic Synechocystis sp.PCC6803 with paralichthys olivaceus growth hormone gene,we conducted experiments through orthogonal tests based on the results of single factor experiments in order to obtain the optimal medium conditions.We used the same method of orthogonal test based on the optimal medium conditions to achieve the best environment factors.The results of our experiment demonstrated that the optimal medium conditions for transgenic strain based on BG-11 medium were determined as follows: The NaNO3,K2HPO4 and Na2CO3 concentrations were 1.25 g/L,0.04 g/L and 0.03 g/L(The rest of the components were the same as BG-11 medium).The optimal culture conditions were temperature 30 ℃,light intensity 3 000 lx and pH 8.0.Under these optimal culture conditions,the biomass of the transgenic alga increases quickly and the specific growth rate of the transgenic Synechocystis sp.PCC6803 reached 20.64.This paper lays an academic foundation for achieving high density and amplificatory culture of transgenic Synechocystis sp.PCC6803.