PROMOTER INFLUENCE ON BACULOVIRUS-MEDIATED GENE-EXPRESSION IN PERMISSIVE AND NONPERMISSIVE INSECT CELL-LINES
PROMOTER INFLUENCE ON BACULOVIRUS-MEDIATED GENE-EXPRESSION IN PERMISSIVE AND NONPERMISSIVE INSECT CELL-LINES
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DOI：
10.1128/jvi.66.12.7397-7405.1992
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发表时间：
1992-12-01
影响因子：
5.4
通讯作者：
MILLER, LK
中科院分区：
文献类型：
--
作者：
MORRIS, TD;MILLER, LK
在一系列昆虫细胞系中检测了病毒启动子和昆虫启动子的活性，这些启动子对加利福尼亚签名杆状病毒核型多角体病毒的复制具有许可性和非许可性。构建重组杆状病毒，将细菌氯霉素乙酰转移酶基因置于病毒复制早期、晚期或极晚期高度活跃的启动子控制之下。在完全受纳细胞中，极晚启动子的表达比晚启动子高2- 3倍，比早期启动子或病毒传播的昆虫启动子高10- 20倍。在不支持病毒子代高效生产的细胞系中，晚期启动子驱动的表达与非常晚期启动子驱动的表达相似或超过。在非受纳昆虫细胞系中，由黑腹果蝇衍生的昆虫启动子驱动的表达高于三种病毒启动子的表达，在果蝇细胞系中表达尤其高。令人惊讶的是，依赖于DNA复制的晚期启动子驱动的表达，在四种非允许系中有三种高于早期启动子驱动的表达。相反，非常晚的启动子驱动的表达在非允许细胞系中非常有限。结果表明，用于驱动外源基因表达的启动子强烈影响了重组杆状病毒感染过程中能够有效支持外源蛋白产生的昆虫细胞范围。
The activities of viral and insect promoters were examined in a range of insect cell lines permissive and nonpermissive for the replication of the baculovirus Autographa californica nuclear polyhedrosis virus. Recombinant baculoviruses were constructed to place the bacterial chloramphenicol acetyltransferase gene under the control of promoters strongly active in the early, late, or very late stages of virus replication. In fully permissive cells, expression from a very late promoter was 2- to 3-fold higher than expression from a late promoter and 10- to 20-fold higher than expression from an early promoter or from a virus-borne insect promoter. In cell lines that do not support the efficient production of viral progeny, late-promoter-driven expression was similar to or surpassed very late promoter-driven expression. In nonpermissive insect cell lines, expression driven by an insect promoter derived from Drosophila melanogaster was higher than expression from the three viral promoters and was especially high in the Drosophila cell line tested. Surprisingly, late-promoter-driven expression, which is dependent on DNA replication, was higher than early-promoter-driven expression in three of four nonpermissive lines. In contrast, very late promoter-driven expression was quite limited in nonpermissive cell lines. The results indicate that the promoter used to drive foreign-gene expression strongly influences the range of insect cells which can efficiently support the production of the foreign protein during infection with recombinant baculoviruses.