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重组逆转录病毒浓缩方法研究
引用本文:王凤阳,扈荣良,邹啸环,池海英,杜卫华,赵君,张守峰,李敏,殷震. 重组逆转录病毒浓缩方法研究[J]. 生物学杂志, 2001, 18(2): 11-13
作者姓名:王凤阳  扈荣良  邹啸环  池海英  杜卫华  赵君  张守峰  李敏  殷震
作者单位:解放军军需大学全军基因工程实验室,长春,130062
基金项目:国家"863"计划资助项目(Z21-04-03)
摘    要:选择含有新霉素磷酸转移酶Ⅱ(Neor)的重组逆转录病毒载体,通过包装细胞进行包装,得到了含有重组逆转录病毒粒子的病毒溶液,该病毒溶液分别采用差速离心和滤膜截留两种方法进行浓缩,浓缩前,后的病毒溶液体外感染靶细胞NIH3T3以测定其滴度。结果显示,差速离心法的浓缩效率要优于滤膜截留法的浓缩效率,其浓缩效率能达到34.5%。

关 键 词:重组逆转录病毒 差速离心 滤膜截留 浓缩方法 载体
文章编号:1008-9632(2001)02-0011-03
修稿时间:2000-10-23

Study on in vitro concentration of recombinant retrovirus
WANG Feng-yang,HU Rong-liang,CHI Hai-ying,DU Wei-hua,ZHAO Jun,ZHANG Shou-feng,LI Min,YIN Zhen. Study on in vitro concentration of recombinant retrovirus[J]. Journal of Biology, 2001, 18(2): 11-13
Authors:WANG Feng-yang  HU Rong-liang  CHI Hai-ying  DU Wei-hua  ZHAO Jun  ZHANG Shou-feng  LI Min  YIN Zhen
Abstract:Recombinant retroviral vectors(pLXSN) carrying Neo r were transfected into PA317 by lipofectin and calcium phosphate coprecipitation.The cell supernatants from G418-resistant PA317 in which there are recombinant retrovirus were concentrated through speed differential centrifugation and membrane filteration,respectively,NIH3T3 cells were infected by the concentrated virus to measure its titer.The results showed that the concentration efficiency of speed differential centrifugation is superior to that of membrane filteration and the concentration efficiency of speed differential centrifugation speed can attain 34.5%.
Keywords:recombinant retrovirus  concentration  speed differential centrifugation  membrane filteration.
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