三甲基月桂基硫酸钠（TLS）在蓝细菌核酸提取中的作用

本文首次把三甲基月桂基硫酸钠(TLS)应用于蓝细菌(蓝藻)总核酸的提取。在用溶菌酶破坏满江红鱼腥藻的细胞壁后,再用TLS分别取代常用法中的各种试剂,即:Triton X-100 Sarkosyl,蛋白酶K,总核酸产率分别是常用法的98.9％,47.3％,103.8％。用TLS取代蛋白酶K处理不同生长期的满江红鱼腥藻,总核酸产率随生长期延长而降低。TLS取代蛋白酶K提取的DNA和常用法提取的DNA,都能被限制性内切酶EcoR I水解。用光学显微镜观察到,满江红鱼腥藻经TLS处理后,丝状体断裂,细胞破裂成碎片。在扫描电子显徽镜下,未经TLS处理的细胞表面光滑平整,而经TLS处理的细胞表面有鳞片状突起、皱褶、凹陷,甚至穿孔。

THE APPLICATION AND FUNCTION OF TRIMETHYLAURYLSODIUM SULFATE IN THE EXTRACTION OF NUCLEIC ACIDS FROM ANABAENA AZOLLAE

In this work trimethylaurylsodium sulfate(TLS) was used as a novel reagent for extraction of total cellular nucleic acids from the cyanobacterium, Anabaena azollae. It is found that TLS can replace Triton X-100 and proteinase K, but can not take the place of Sarkosyl in standard method. Compared with the standard method, the productivity of total nucleic acids from the cells was 98.9%, 103.8% and 47.3% respectively, When TLS replaced Triton X-100, proteinase K or Sarkosyl. The effect of TLS varied with the growth phase of the cyanobacterium. The test of restriction endonuclease digestion by EcoR I indicated that DNA extracted and purified by the method of TLS replacing proteinase K was sensitive to cleavage. After treated with lysozyme, the filaments broke down into singular cells. Then after treated with TLS, the debris of broken cells were observed under an optical microscope. Also scale-like degradation and perforation were found on the wrinkled cell surface of Anabaena azollae from observations using SEM.