摘要
微型腺病毒(mAd)是去除所有编码基因,仅保留两侧反向末端重复序列(ITR)及包装信号(Ψ)的新型腺病毒载体.它具有包装容量大,免疫原性小等优点.在大量扩增时,需与缺失E1区及包装信号的辅助腺病毒共转染入包装细胞(293细胞)内,CsCl超离心纯化,将二者分开.再通过琼脂糖覆盖挑斑、在包装细胞内大量扩增、CsCl超离心纯化等步骤,最终得到较为纯净的mAd.但因mAd结构不稳定,易与辅助病毒发生同源重组,在CsCl超离心时不能完全与辅助病毒分开,导致外源基因表达逐步下降.
Mini-Adenovirus (mAd) is a novel adenoviral vector that contains only the viral inverted terminal repeat se- quences (ITR), packaging signal and cis-acting elements required for viral DNA replication and packaging. MAd vec- tor has larger capacity and lower immunogenicity, which showed its proprietary prospect in the field of gene therapy. For amplification of mAd vector, the cells must be co-transfected with mAd and helper Ad that deleted El region and packaging signal for packaging cells. Through plaque assay, amplification and propagation , the density of purified mAd was 2.3× 10^(11) mL^(-1) However, mAd genome was found unstable and with 1% contamination of helper Ad. The am- plification and purification of mAd were discussed.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
2001年第5期525-529,共5页
Journal of Fudan University:Natural Science
基金
国家"863"项目(863-Z20-02-01)
国家自然科学基金(39880019)
国家教育部优秀青年教师基金
教育部