作　　者： ; ; ;

机构地区： 中国科学院武汉植物研究所武汉植物园

出　　处： 《武汉植物学研究》 2003年第4期295-300,共6页

摘　　要： 四爿藻具有坚硬的囊壳和特殊的细胞壁组成,细胞不易破碎,且含有丰富的糖蛋白,易对DNA造成污染,使其基因组DNA提取较为困难、纯度不高。研究对比传统的CTAB法与高盐低pH法提取四爿藻DNA,高盐低pH法快速经济,得到的基因组DNA纯度较高,是一种行之有效的四爿藻基因组DNA提取方法。该法通过改变抽提介质,提高细胞破碎效率,减少抽提次数,有效去除污染,提取的基因组DNA不仅可以成功进行nrDNA转录间隔区(ITS)扩增,而且扩增产物适合于进行测序分析,这为其它淡水和海洋单细胞及多细胞藻类基因组DNA的提取也提供了借鉴。 Tetraselmis,a unicellular green algae located uniquely in phytophylogeny,is a new important model organism for the study of the origin and evolution of green plants.The genomic DNAs of Tetraselmis are difficult to be isolated because of its hard theca and unique cell wall components,as well as its richness in glycoprotein easy to impurify the genomic DNA.We reported here that an improved low pH extraction matrix with high salts originally developed for higher plant was successfully to obtain high quality of genomic DNA of Tetraselmis.Compared to the traditional CTAB extraction,this approach is very fast and economical just by changing extraction medium so that the efficiency of Tetraselmis cell breakage was greatly improved,and the pollution of proteins released during DNA extraction were drastically reduced as well.The fragments of Tetraselmis nuclear ribosome internal transcribed DNA spacers (nrDNA ITS) were PCRamplified successfully from this minipreparations of the genomic DNA,but also amenable to nrDNA ITS sequencing.Due to its application to some higher plants whose genomic DNAs are hard to be isolated,this improved method of low pH extraction matrix with high salts we described here for the marine unicellular microalga Tetraselmis could be right for the minipreparation of genomic DNA of other unicellular or multicellular fresh and marine algae.

关 键 词： 四爿藻 基因组 提取 高盐低 法 法

领　　域： [生物学] [生物学]