摘要
目的 :探讨磷酸钙沉淀法同源重组穿梭质粒 p Ad CMV- tk与腺病毒基因组改装质粒 p JM17的可行性。方法 :用磷酸钙沉淀法将质粒 p Ad CMV- tk与质粒 p JM17共转染至 2 93细胞 ,获得复制缺陷的重组腺病毒 Ad-CMV- tk,将其在 2 93细胞中扩增 ,氯化铯密度梯度离心法纯化扩增病毒 ,空斑实验测定病毒滴度。结果 :质粒 Ad-CMV- tk与质粒 p JM17在 2 93细胞中同源重组后 7～ 10天出现完全的细胞病变态反应。纯化后病毒滴度为 3× 10 1 0pfu/ ml。结论 :磷酸钙沉淀法能成功获得同源重组病毒 。
Objective:To evaluate the feasibility of recombining plasimd pAdCMV-tk and pJM17 by calcium phosphate deposit.Methods:pAdCMV-tk and pJM17 plasmid was transferred into 293 packaged cell by calcium phosphate deposit, then amplified in 293 cell , after purified ,plaque assay was used to determine the titer of AdCMV-tk. Results: 7～10 days after recombinant,in 293 cell Cytopathic Effect(CPE) occurred,after purified, AdCMV-tk had high titers of virus( 3×10 10 pfu/ml). Conclusions: Calcium phosphate deposit method can be successfully used to recombine adeno-virus vector and after purified can obtain high titer virus.
出处
《南通医学院学报》
2002年第4期397-399,共3页
ACTA Academiae Medicinae Nantong
关键词
腺病毒
重组
磷酸钙沉淀法
细胞病变态反应
adeno-virus
recombine
calcium phosphate deposit
cytopathic effect
