构建含微型oriP的基于爱泼斯坦-巴尔病毒的表达载体。

Construction of Epstein-Barr virus-based expression vector containing mini-oriP.

作者信息

Tanaka J, Miwa Y, Miyoshi K, Ueno A, Inoue H

机构信息

Department of Biochemistry, School of Dentistry, University of Tokushima, 3-18-15 Kuramoto, Tokushima, 770-8504, Japan.

出版信息

Biochem Biophys Res Commun. 1999 Nov 2;264(3):938-43. doi: 10.1006/bbrc.1999.1617.

DOI:10.1006/bbrc.1999.1617

PMID:10544034

Abstract

Epstein-Barr virus (EBV)-based vectors are extrachromosomal vectors carrying a replicational origin, oriP (about 2200 bp) and a replication initiation factor (EBNA-1) which are sufficient for autonomous replication. Because one disadvantage of these vectors is their large sizes, we examined the effect of partial deletion of oriP on the effectiveness of the EBV-based vectors, using an enhanced green fluorescent protein (EGFP) as a reporter to monitor gene expression. Results indicated that 954 bp-deleted mini-oriP is useful in primate cells since the vector showed high efficiency of stable transfection, a high ratio of EGFP-positive cells, and high recovery of intact plasmid DNA from transfected cells.

摘要

基于爱泼斯坦-巴尔病毒（EBV）的载体是携带复制起点oriP（约2200碱基对）和复制起始因子（EBNA-1）的染色体外载体，这些对于自主复制而言已足够。由于这些载体的一个缺点是它们的尺寸较大，我们使用增强型绿色荧光蛋白（EGFP）作为报告基因来监测基因表达，研究了oriP部分缺失对基于EBV的载体有效性的影响。结果表明，缺失954碱基对的微型oriP在灵长类细胞中是有用的，因为该载体显示出稳定转染效率高、EGFP阳性细胞比例高以及从转染细胞中回收完整质粒DNA的回收率高。