%0 Journal Article
%T 聚乙烯亚胺提升慢病毒滴度的研究
%A 张宏鹏
%A 唐小川
%A 徐世永
%A 陈清
%A 刘红林
%J 南京农业大学学报
%D 2016
%R 10.7685/jnau.201507029
%X [目的] 本文旨在解决传统慢病毒生产方法中存在的实际操作复杂或成本高等问题,提高慢病毒生产效率。[方法] 使用了以聚乙烯亚胺(PEI)为转染介质,以聚乙二醇6000(PEG6000)为离心浓缩介质的慢病毒生产方法,并对浓缩后的慢病毒进行滴度测定。还对可能影响慢病毒滴度的几个关键因素――转染初始细胞数、N/P值、转染质粒DNA量、佐剂的选择等的作用进行了分析。[结果] 在使用15 cm培养皿生产慢病毒的条件下,转染质粒DNA总量10.5 μg,N/P=20,PEI浓储液pH值为7.0~9.0,转染时293T细胞数(0.5~2)×107时,可以获得较好的慢病毒滴度。[结论] PEI作为一种新的转染试剂,可以替代磷酸钙和脂质体用于高滴度慢病毒的生产。</br>[Objectives] The paper aims to solve the problems of the actual operation in the traditional lentivirus production method, for example, its process is very complicated and the experimental cost is too high.[Methods] The polyethylenimine(PEI)was used as the medium of the transfection and polyethylene glycol 6000(PEG6000)was used as the centrifugal concentrated medium in the lentivirus production process. In addition, the effect of several possible key factors that could influence the titer of the virus was analyzed.[Results] We obtained excellent lentivirus using suitable conditions, which include:total amount of plasmid DNA about 10.5 μg, N/P=20, PEI concentrated liquid storage pH changing from 7.0 to 9.0, transfected 293T cells number changing from 0.5×107 to 2×107 in a 15 cm plate.[Conclusions] PEI can replace calcium phosphate and liposome for the production of high titer lentivirus
%K 慢病毒生产
%K 聚乙烯亚胺
%K 载体构建
%K 滴度检测
%K 转染试剂&lt
%K /br&gt
%K lentivirus production
%K polyethylenimine(PEI)
%K vector construction
%K titer detection
%K transfection reagent
%U http://nauxb.njau.edu.cn/oa/darticle.aspx?type=view&id=201603015