Baculovirus-mediated gene transfer into mammalian cells
Baculovirus-mediated gene transfer into mammalian cells
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DOI：
10.1073/pnas.93.6.2348
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发表时间：
1996-03-19
影响因子：
11.1
通讯作者：
Bucher, NLR
中科院分区：
文献类型：
--
作者：
Boyce, FM;Bucher, NLR
本文报道了利用杆状病毒Arctographa californica多核多角体病毒（AcMNPV）作为载体将基因导入哺乳动物细胞，制备了一种带有大肠杆菌lacZ报告基因的改良AcMNPV病毒，该报告基因受劳斯肉瘤病毒启动子和哺乳动物RNA加工信号的控制，然后用这种改良的AcMNPV病毒感染多种哺乳动物细胞系，在感染人肝细胞系HepG 2后，>25%的细胞显示高水平的转导基因表达，在暴露于病毒后，在大鼠肝细胞的原代培养物中超过70%的细胞显示β-半乳糖苷酶表达，来自其他组织的细胞系在暴露于病毒后显示较少或不表达lacZ，在不太敏感的细胞中对表达的阻断似乎不是由被靶细胞内化的能力引起的，而是由病毒进入后的事件引起的。在HepG 2细胞中，lacZ表达的开始发生在感染后6小时内，并在感染后12-24小时达到峰值。AcMNPV能够携带大的（> 1.5kb）插入片段，是肝细胞原代培养的高效基因传递载体，AcMNPV可能是肝细胞遗传操作的有用载体。
This paper describes the use of the baculovirus Arctographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a vector for gene delivery into mammalian cells, A modified AcMNPV virus was prepared that carried the Escherichia coli lacZ reporter gene under control of the Rous sarcoma virus promoter and mammalian RNA processing signals, This modified baculovirus was then used to infect a variety of mammalian cell lines, After infection of the human liver cell lines HepG2, >25% of the cells showed high-level expression of the transduced gene, Over 70% of the cells in primary cultures of rat hepatocytes showed expression of beta-galactosidase after exposure to the virus, Cell lines from other tissues showed less or no expression of lacZ after exposure to the virus, The block to expression in less susceptible cells does not appear to result from the ability to be internalized by the target cell but rather by events subsequent to viral entry, The onset of lacZ expression occurred within 6 hr of infection in HepG2 cells and peaked 12-24 hr postinfection, Because AcMNPV is able to replicate only in insect hosts, is able to carry large (>1.5 kb) inserts, and is a highly effective gene delivery vehicle for primary cultures of hepatocytes, AcMNPV may be a useful vector for genetic manipulation of liver cells.