以雨生红球藻(Haematococcusplauvialis)34-1n和192.80为研究材料,采用细胞计数法对它们的生长特性进行分析。在BBM培养基中加入不同浓度的Zeomycin、腐草霉素和巴龙霉素等常用筛选试剂,通过细胞计数和显微镜观察等方法对各抗生素对藻细胞的作用进行分析,结果表明,雨生红球藻34—1n更适合作为遗传转化的受体藻株;进一步分析表明,它对巴龙霉素会产生回复突变,而对Zeomycin敏感,8μg/mLZeomycin为最适的筛选试剂浓度。因此,Zeomycin相应的抗性基因ble可作为雨生红球藻遗传转化的筛选标记基因。
Haematococcus plauvidis 34-1n and 192.80 were employed in this research. In order to analyze their growth characters, counting-cell-method and microscope were used. After adding different concentration of zeomycin, phleomycin and paromomycin to BBM medium, cells were counted and observed using microscope. Results showed that H. plauvialis 31-1n was more suitable for genetic transformation than H. plauvialis 192.80. Further studies also indicated that H. plauvialis 31-ln was highly sensitive to zeomycin, while not sensitive to paromomycin. The concentration of 8 μg/mL zeomycin might be a suitable concentration when building up transformation system in H. plauvi(dis. Hence, it suggested that zeomycin could be suitable agent and ble gene could be a seleetable marker for genetic transformation in H. plauvialis.