Citation: PENG Yu, LI Shu, LI Chao-yang, WANG Jing-jing, TIAN Bo, GUO De-yin*. Interaction between Nucleocapsid Protein and the Leader the Sequence of SARS-CoV RNA .VIROLOGICA SINICA, 2005, 20(5) : 490-493.

Interaction between Nucleocapsid Protein and the Leader the Sequence of SARS-CoV RNA

  • Available online: 20 October 2005
  • SARS coronavirus(SARS-CoV) is a novel human coronavirus and was responsible for SARS outbreak in several regions of the world in 2003.The genome of SARS-CoV is about 30,000 nucleotides in length and sequence analysis revealed that nucleotides 1-72 contain a RNA leader sequence preceding an untranslated region(UTR).In order to confirm the interaction between the N protein and the RNA leader sequence,the N gene was cloned,sequenced and the protein was expressed in Escherichia coli as a fusion protein with His-tag.The recombination N protein was purified by Ni-NTA affinity column and G-75 column.Meanwhile,the DIG labeled RNA leader sequences were synthesized by in vitro transcription.RNA-Protein interaction was detected by Northwestern RNA blot assay.The results indicated that a specific interaction takes place between the N protein and RNA leader sequence.Our results provide some insights into the understanding of assembly of the SARS-CoV particles,which is an essential step for viral replication cycle.

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    Interaction between Nucleocapsid Protein and the Leader the Sequence of SARS-CoV RNA

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    Abstract: SARS coronavirus(SARS-CoV) is a novel human coronavirus and was responsible for SARS outbreak in several regions of the world in 2003.The genome of SARS-CoV is about 30,000 nucleotides in length and sequence analysis revealed that nucleotides 1-72 contain a RNA leader sequence preceding an untranslated region(UTR).In order to confirm the interaction between the N protein and the RNA leader sequence,the N gene was cloned,sequenced and the protein was expressed in Escherichia coli as a fusion protein with His-tag.The recombination N protein was purified by Ni-NTA affinity column and G-75 column.Meanwhile,the DIG labeled RNA leader sequences were synthesized by in vitro transcription.RNA-Protein interaction was detected by Northwestern RNA blot assay.The results indicated that a specific interaction takes place between the N protein and RNA leader sequence.Our results provide some insights into the understanding of assembly of the SARS-CoV particles,which is an essential step for viral replication cycle.

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