用甲基化修饰法抑制钝顶螺旋藻遗传转化中限制性内切酶对外源质粒的降解  

Methylating the foreign plasmids on Spirulina/Arthrospira platensis transformation to resist the restriction endonuclease digestion

作　　者：王迪[1] 卢永忠[1] 张学成[1] 茅云翔[1] 隋正红[1] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《海洋科学》2007年第3期1-4,8,共5页Marine Sciences

基　　金：国家自然科学基金资助项目(30471317)

摘　　要：胞内限制性内切酶降解外源DNA,是钝顶螺旋藻/节旋藻(Spirulina/Arthrospira platensis)遗传转化的难点之一。通过用EDTA螯合Mg2+抑制限制性内切酶活性的方法,对外源质粒进行了甲基化修饰。结果表明,修饰后的外源质粒可抵抗限制性内切酶3h的降解,有利于钝顶螺旋藻的遗传转化。The active restriction endonuclease of Spirulina/Arthrospira platensis is believed to be the most serious barrier of the foreign gene transformation.With EDTA chelating Mg^2＋,we methylate the foreign plasmids by using the crude extracts of cells. Results show that the foreign plasmids can resist the digestion of restriction endonuclease for three hours,and the methylated plasmids are better than tmmethylated plasmids for transformation.

关 键 词：钝顶螺旋藻(Spirulina/Arthrospira platensis) 限制修饰系统 甲基化酶 

分 类 号：Q291[生物学—细胞生物学]