转基因螺旋藻的光生物反应器培养及鉴定

High-density Cultures and Identification of the Transgenic Spirulina platensis

以本实验室构建的转胸腺素基因螺旋藻为藻种，利用中科院过程工程研究所研制的PhR-L20C光生物反应器，根据该反应器的各项性能指标和参数条件，采用本实验室优化过的培养条件，对转化藻进行高密度培养，初始pH值约为9.0，光照强度平均约为10000Lx，培养温度28-30℃，一个培养收获周期为10d，平均生长速率为0.3OD560·d，10d后藻细胞OD560达3.0，最终藻细胞密度可达到1.3g·L。（干重）；高密度培养后的转化藻经PCR鉴定，目的基因能够稳定地整合在螺旋藻染色体上，经Western-blot证明外源基因在螺旋藻中获得高效表达，且在光生物反应器中经过30d的无抗生素G418的高密度培养后，外源基因能够稳定的遗传表达。

The transgenic Spirulina platensis constructed in our lab was cultured in a PhR - L20C photobioreactor designed by the Institute of Process Engineering, Chinese Academy of Science. The conditions were optimized and applied for the high-density cultivation within the parameters of this photobioreactor. The initial pH was about 9.0, light intensity 10000 Lx, and the temperature 28-30℃. The result showed that the average optical density （ODs50） reached 3.0 after 10 days with a growth rate of 0.30D reading per day. The final average dry weight yield of S. platensis was 1.3 g· L^-1. After cultivation for 30 days without the aid of the antibiotic, G418, in the photobioreac-tor, the target gene was positively detected by means of PCR on the genome of S. platensis. In addition, the West-ern-blot result proved the foreign gene was effectively expressed in the transgenic S. platensis.