提取条斑紫菜高纯度总DNA及其质粒状DNA的新方法  

New Methods for Extraction of Plasmid-like DNA and High-Quality Total DNA from Porphyra yezoensis

作　　者：郭宝太[1] 夏连胜[1] 姜国勇[1] 毕玉平[2] 戴继勋[3] 

机构地区：[1]莱阳农学院农学系,山东莱阳265200 [2]山东省农科院生物技术中心 [3]青岛海洋大学海洋生命学院

出　　处：《莱阳农学院学报》2002年第1期3-6,共4页Journal of Laiyang Agricultural College

摘　　要：本文报道提取条斑紫菜高纯度总DNA及其质粒状DNA的新方法。先用海螺酶处理紫菜叶状体制备紫菜体细胞 ,然后用SDS -蛋白酶K裂解细胞提取总DNA ,再用WizardDNAclean -up树脂对其纯化 ,经纯化后的总DNA能被EcoRI、PstI等限制酶完全酶解 ,并在酶切图谱上形成明显的DNA带型。当用异硫氰酸胍—十二烷基肌氨酸钠裂解紫菜细胞时 ,在总DNA提取物中直接发现有一条质粒状DNA带 。Somatic cells were prepared from sea snail enzyme digests of Porphyra yezoensis thalli. Using SDS-Proteinase K as extraction solution, total DNA was isolated from the somatic cells. The crude extracts of total DNA were purified with Wizard DNA clean-up resin, and the resulting DNA was of sufficient quality for digestion of restriction endonuclease. DNA bands were clearly observed in the restriction patterns of EcoRI, PstI respectively. The presence of DNA bands in the restriction pattern of total DNA indicated that the genome of Porphyra yezoensis may be small. Unexpectedly, using guanidinium isothiocyanate and sarcosyl as extraction solution, a plasmid-like DNA band (2.3 Kb) was directly found in the isolated total DNA of Porphyra yezoensis. A very simple and convenient method for isolation of plasmid-like DNA has been established.

关 键 词：条斑紫菜 体细胞 总DNA 限制性酶切 质粒状DNA 

分 类 号：Q959.292.1[生物学—动物学] Q523