适合膜片钳电流测定的金铁锁原生质体制备技术  

Isolation of protoplast from the Psammosilene tunicoides suitable for the whole-cell patch-clamp experiment

作　　者：赵欣[1] 李岩[1] 王丽爽[1] 张宗申[1] 

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034

出　　处：《大连工业大学学报》2012年第1期24-27,共4页Journal of Dalian Polytechnic University

基　　金：国家自然科学基金资助项目(31070164)

摘　　要：以金铁锁愈伤组织为材料,利用传统酶解法和快速酶解法,对金铁锁原生质体的制备分离技术进行比较和研究,记录全细胞电流,获得高质量的原生质体材料。结果表明,采用传统酶解法,2%纤维素酶R-10+1%果胶酶Y-23的混合酶液酶解7h,可以获得大量原生质体;采用快速酶解法,1.2%纤维素酶RS+0.5%果胶酶Y-23的混合酶液酶解15min,然后分别加入300μL、1mL基本介质稀释后,在400mmol/kg渗透压下可以获得高质量的原生质体。全细胞电流测定表明,快速酶解法制得的原生质体更适于作为膜片钳实验的材料。Callus of Psammosilene tunicoides was used to study the effects of conditions on its protoplast preparation by taking the traditional method and the fast enzymolysis method with record the current of entire cell to gain the suitable protoplast.The results showed that massive protoplasts were available in the traditional method using 2% cellulase R-10+1% Pectolyase Y-23 to treat the callus for 7 h;in the fast enzymolysis method,the optimized treatment was 1.2% cellulase RS+0.5% pectolyase RS for 15 min,and then diluted by 300 μL,1 mL basic solution under 400 mmol/kg osmotic pressure.It's proved that the protoplasts by the fast enzymolysis method were more suitable for patch-clamp experiment compared with that by traditional method.

关 键 词：金铁锁 原生质体 膜片钳 

分 类 号：S681.5[农业科学—观赏园艺]