摘要
目的改进AdEasy系统,使之更简便、高效、快捷。方法pAdtrack经酶切线性化后,转化经氯化钙处理的感受态AdEasier-1细菌,获得重组腺病毒质粒。最后将线性化的重组腺病毒质粒转染293细胞包装获得重组腺病毒。荧光显微镜下细胞中重组腺病毒观察绿色荧光蛋白的表达。结果应用氯化钙法由pAdtrack转化AdEasier-1细菌,可获得极高比例的(20/20)阳性重组腺病毒克隆。结论应用改进后的AdEasy系统构建腺病毒相比于传统AdEasy系统,方法更简便、成功率更高、实验周期更短、对实验室的条件要求低。
To simplify AdEasy system for better efficiency, we atte mp ted some modifications on the original system. Specifically, the PmeⅠ-li nearized plasmid pAdtrack was transformed into competent AdEasier-1 cells prepar ed from trea-tment with CaCl 2 . The DNA contained in the identified recombinant plasmid was digested with PacⅠand transfected into 293 cells to package the adenovirus, followed by iden tification of the recombinant adenovirus by means of observation of green fluor escence protein expression under fluorescentce microscope. Chemical trans formation of the linearized plasmid into AdEasier-1 cells resulted in very high success rate (20/20) for producing positive recombinant adenovirus clones, confirming the efficiency of the modified AdEasy system in constructing recomb inant adenovirus.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第5期501-503,共3页
Journal of First Military Medical University
基金
广东省自然科学基金资助项目(013072)
国家863计划项目(2001AA217171)~~
