摘要:
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10 000 Lx,培养温度28~30℃,一个培养收获周期为10 d,平均生长速率为0.3 OD560·d-1,10 d后藻细胞OD560达3.0,最终藻细胞密度可达到1.3 g·L-1(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30 d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。
Abstract:
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(OD560) reached 3.0 after 10 days with a growth rate of 0.3 OD 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 photobioreactor,the target gene was positively detected by means of PCR on the genome of S.platensis.In addition,the Western-blot result proved the foreign gene was effectively expressed in the transgenic S.platensis.
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