透析浓缩法提取与纯化重组腺病毒DNA  

INCREASE THE OUTPUT OF EXTRACTION AND PURIFICATION OF RECOMBINED ADENOVIRUS DNA FROM MEDIUM WITH DIALYSIS

作　　者：王国强[1] 沈岳[1] 李芳[1] 廖维宏[1] 

机构地区：[1]第三军医大学野战外科研究所第三研究室,重庆400042

出　　处：《中国神经科学杂志》1999年第4期331-333,共3页

基　　金：国家自然科学基金!资助项目(39670740)

摘　　要：从重组腺病毒转染人胚肾细胞的培养液中提取及纯化病毒ＤＮＡ。将培养液装入透析袋，袋外用聚乙二醇包埋以吸附培养液中的水分直到袋内液体浓缩至所需量。取出浓缩液，用裂解液裂解病毒，然后进行酚／氯仿抽提，透析去杂质及乙醇沉淀。结果表明，从大量培养液中提出的重组腺病毒ＤＮＡ浓度高（４１．３６ μｇ／μｌ±１．６７ μｇ／μｌ）、纯度高（紫外吸收比值２６０ ｎｍ ／２８０ ｎｍ ＝ １．８），可直接用于酶切反应及ＤＮＡTo extract and purify recombined adenovirus DNA from culture medium,the medium was transferred into a dialysis bag when extensive cell death was observed.The bag was packaged with polyethylene glycol(PEG)which would absorb most of water within the bag, then drew out the concentrated liquid from the bag into a tube and added lysis buffer.Adenovirus DNA was extracted by phenol/chloroform and purified through dialysis in NaCl/sodium citrate followed by precipitation with ethanol.The result showed that both the concentration(41 36 μg/μl±1 67 μg/μl) and the purity (ultraviolet absorbency of 260 nm/280 nm=1 8) of the extracted DNA were very high.The extracted DNA can be directly used in the procedures of endonuclease reaction and DNA probe preparation.

关 键 词：DNA 聚乙二醇 重组腺病毒 透析浓缩法 

分 类 号：R394[医药卫生—医学遗传学] R373.9[医药卫生—基础医学]