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Studies of Coronavirus DI RNA Replication Using In Vitro Constructed DI cDNA Clones

  • Shinji Makino
  • Michael M. C. Lai
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 276)

Abstract

Sequence analysis of an intracellular defective-interfering (DI) RNA, DIssE, of mouse hepatitis virus (MHV) revealed that it is composed of three noncontiguous genomic regions, representing the first 864 nucleotides of the 5′-end, an internal 748 nucleotides of the polymerase gene, and 601 nucleotides from the 3′-end of the parental MHV genome. DIssE had three base substitutions within the leader sequence and also a deletion of nine nucleotides located at the junction of the leader and the remaining genomic sequence. A system was developed for generating DI RNAs to study the mechanism of MHV RNA replication. A cDNA copy of DIssE RNA was placed downstream of T7 RNA polymerase promoter to generate DI RNAs capable of extremely efficient replication in the presence of a helper virus. We demonstrated that, in the DI RNA-transfected cells, the leader sequence of these DI RNAs was switched to that of the helper virus during one round of replication. This high-frequency leader sequence exchange was not observed if a nine-nucleotide stretch at the junction between the leader and the remaining DI sequence was deleted. This observation suggests that a free leader RNA is utilized for the replication of MHV RNA.

Keywords

Leader Sequence Helper Virus Mouse Hepatitis Virus Packaging Signal Murine Coronavirus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1990

Authors and Affiliations

  • Shinji Makino
    • 1
  • Michael M. C. Lai
    • 1
  1. 1.Department of MicrobiologyUniversity of Southern California, School of MedicineLos AngelesUSA

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