紫球藻(Porphyridium cruentum)原生质体分离研究

Study on the Protoplasts Formation from Porphyridium Cruentum

收集对数生长期的紫球藻细胞 ,经 1%氯化苄预处理后 ,用 1 8%纤维素酶和 1 2 %果胶酶组成的混合水解酶液 (含 0 6mol/LNaCl,pH7 2 ) ,在 2 8℃下缓慢振荡酶水解细胞壁 ,4h后原生质体分离率比未预处理组提高 74% ,在最佳分离条件下的原生质体分离率高达 5 6 4% ,基本能满足原生质体融合的需求。

The cells of Porphyridium cruentum cultured 4 days were pretreated with 1% benzyl chloride for 30 minutes , then moved into mixing enzyme solution of 1 8% cellulase and 1 2% pectinase (0 5 mol/L NaCl, pH 7 2) , and shaken gently (50 r/min)under 28℃ The protoplasts release rate amounts to 74% higher than non pretreated ones 4hr latter Under the best circumstance , the protoplasts release rate can be as high as 56 4%, which can be used on protoplast fusion