Xiong C, Levis R, Shen P, Schlesinger S, Rice C M, Huang H V
Department of Microbiology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
Science. 1989 Mar 3;243(4895):1188-91. doi: 10.1126/science.2922607.
Sindbis virus, an enveloped virus with a single-stranded RNA genome, was engineered to express a bacterial protein, chloramphenicol acetyltransferase (CAT), in cultured insect, avian, and mammalian cells. The vectors were self-replicating and gene expression was efficient and rapid; up to 10(8) CAT polypeptides were produced per infected cell in 16 to 20 hours. CAT expression could be made temperature-sensitive by means of a derivative that incorporated a temperature-sensitive mutation in viral RNA synthesis. Vector genomic RNAs were packaged into infectious particles when Sindbis helper virus was used to supply virion structural proteins. The vector RNAs were stable to at least seven cycles of infection. The expression of CAT increased about 10(3)-fold, despite a 10(15)-fold dilution during the passaging. Sindbis virus vectors should prove useful for expressing large quantities of gene products in a variety of animal cells.
辛德毕斯病毒是一种具有单链RNA基因组的包膜病毒，经过改造后可在培养的昆虫、禽类和哺乳动物细胞中表达一种细菌蛋白——氯霉素乙酰转移酶（CAT）。这些载体能够自我复制，基因表达高效且迅速；在16至20小时内，每个受感染细胞可产生多达10⁸个CAT多肽。通过在病毒RNA合成中引入温度敏感突变的衍生物，可使CAT表达呈现温度敏感性。当使用辛德毕斯辅助病毒来提供病毒粒子结构蛋白时，载体基因组RNA会被包装成感染性颗粒。载体RNA在至少七个感染周期内保持稳定。尽管在传代过程中稀释了10¹⁵倍，但CAT的表达仍增加了约10³倍。辛德毕斯病毒载体应可证明在多种动物细胞中表达大量基因产物方面很有用。
