Jalanko A, Kallio A, Ulmanen I
Orion Genetic Engineering Laboratory, Helsinki, Finland.
Arch Virol. 1988;103(3-4):157-66. doi: 10.1007/BF01311089.
We have characterized the properties of an Epstein-Barr virus vector (EBV-CMV) and compared its expression potential with a respective integrating vector (CMV). These vectors were used to express chloramphenicol acetyltransferase (CAT) gene in human HeLa, 293, monkey CV-1, dog MDCK, and hamster R 1610 cells. The EBV-CMV-cat DNA replicates extrachromosomally in HeLa, 293 and CV-1 cells, where also high expression of CAT gene was observed. The EBV-CMV vector integrated in MDCK and R 1610 cells and the CMV vector integrated in all cells tested. Integration yielded mostly clones with low CAT expression. In all cell lines, except HeLa cells, the existence of the extrachromosomal but not the integrated vector DNA is strictly dependent on the Hygromycin B selection pressure. The extrachromosomal state of the EBV vector is a prerequisite for good expression particularly in human and monkey cells.
我们已对爱泼斯坦-巴尔病毒载体（EBV-CMV）的特性进行了表征，并将其表达潜能与相应的整合载体（CMV）进行了比较。这些载体用于在人HeLa细胞、293细胞、猴CV-1细胞、犬MDCK细胞和仓鼠R 1610细胞中表达氯霉素乙酰转移酶（CAT）基因。EBV-CMV-cat DNA在HeLa细胞、293细胞和CV-1细胞中进行染色体外复制，在这些细胞中也观察到了CAT基因的高表达。EBV-CMV载体整合到MDCK细胞和R 1610细胞中，而CMV载体整合到所有测试细胞中。整合产生的大多是CAT表达水平低的克隆。在所有细胞系中，除HeLa细胞外，染色体外而非整合的载体DNA的存在严格依赖于潮霉素B选择压力。EBV载体的染色体外状态是良好表达的先决条件，尤其是在人和猴细胞中。
