Sindbis virus: an efficient, broad host range vector for gene expression in animal cells.
Sindbis virus: an efficient, broad host range vector for gene expression in animal cells.
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DOI：
10.1126/science.2922607
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发表时间：
1989-03
期刊：
影响因子：
56.9
通讯作者：
C. Xiong;R. Levis;P. Shen;S. Schlesinger;C. Rice;Henry Huang
中科院分区：
文献类型：
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作者：
C. Xiong;R. Levis;P. Shen;S. Schlesinger;C. Rice;Henry Huang
辛德比斯病毒是一种具有单链RNA基因组的包膜病毒，它被改造成在昆虫、禽类和哺乳动物细胞中表达细菌蛋白氯霉素乙酰转移酶(CAT)。该载体具有自我复制能力，基因表达效率高，表达速度快；每个感染细胞在16~20小时内可产生多达10(8)个CAT多肽。通过在病毒RNA合成中加入温度敏感突变的衍生物，可以使CAT的表达对温度敏感。当Sindbis Helper病毒提供病毒结构蛋白时，载体基因组RNA被包装成具有感染性的颗粒。载体RNA在至少七个感染周期中都是稳定的。尽管在传代过程中CAT的表达稀释了10(15)倍，但CAT的表达增加了约10(3)倍。辛德比斯病毒载体应该被证明在各种动物细胞中表达大量基因产物是有用的。
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.