Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis

被引：176

作者：

Almazan, Fernando ^{[1
]}

DeDiego, Marta L. ^{[1
]}

Galan, Carmen ^{[1
]}

Escors, David ^{[1
]}

Alvarez, Enrique ^{[1
]}

Ortego, Javier ^{[1
]}

Sola, Isabel ^{[1
]}

Zuniga, Sonia ^{[1
]}

Alonso, Sara ^{[1
]}

Moreno, Jose L. ^{[1
]}

Nogales, Aitor ^{[1
]}

Capiscol, Carmen ^{[1
]}

Enjuanes, Luis ^{[1
]}

机构：

[1] CSIC, Ctr Nacl Biotecnol, Dept Mol & Cell Biol, E-28049 Madrid, Spain

来源：

JOURNAL OF VIROLOGY | 2006年 / 80卷 / 21期

关键词：

D O I：

10.1128/JVI.00385-06

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The engineering of a full-length infectious cDNA clone and a functional replicon of the severe acute respiratory syndrome coronavirus (SARS-CoV) Urbani strain as bacterial artificial chromosomes (BACs) is described in this study. In this system, the viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and further amplified in the cytoplasm by the viral replicase. Both the infectious clone and the replicon were fully stable in Escherichia coli. Using the SARS-CoV replicon, we have shown that the recently described RNA-processing enzymes exoribonuclease, endoribonuclease, and 2'-O-ribose methyltransferase were essential for efficient coronavirus RNA synthesis. The SARS reverse genetic system developed as a BAC constitutes a useful tool for the study of fundamental viral processes and also for developing genetically defined vaccines.

引用

页码：10900 / 10906

页数：7

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