雨生红球藻农杆菌转化体系的建立

Establishment of transformation system of Agrobacterium in Haematococcus pluvialis

文章以植物表达载体pBI121、农杆菌EHA105为体系,建立了农杆菌介导的雨生红球藻转化方法,通过筛选羧苄青霉素、G418等抗生素对雨生红球藻生长的影响,确定了以200μg/L G418和500mg/L羧苄青霉素为筛选体系。转化的雨生红球藻以CaMV 35S和来源于番茄的八氢番茄红素脱氢酶基因的启动子(PDS启动子)成功表达了报告基因GFP和YFP,拓宽了pBI121载体的应用范围,为雨生红球藻的转化提供了一个新的遗传转化途径。番茄来源的PDS启动子能够启动报告基因的表达,表明植物源的启动子能够在雨生红球藻中表达,为植物源的启动子验证及基因瞬时表达提供了一个新的方法。

The Agrobacterium‐mediated transformation method of Haematococcus pluvialis（H .pluvialis） was established by using plant expression vector pBI 121 and A grobacterium tume f aciens EHA105 .The optimum concentration of G418 and carbenicillin was 200μg/L and 500 mg/L ,respectively ,by screening for the effect of carbenicillin and G418 on alge H .pluvialis growth .Reporter gene GFP or YFP driven by CaMV 35S or tomato PDS promoter ,respectively ,was successfully expressed in H .pluvialis ,suggesting that the applica‐tion of pBI121 vector was broadened ,and thereby providing a new way for the genetic transformation of H . p luv ialis .Tomato‐derived PDS promoter is able to activate reporter gene expression ,indicating that botanical promoter can be expressed in H .pluvialis ,which provides a new method for the verification of promoters from plants and transient gene expression .