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| Expression and Purification of C-terminal of Arenavirus Polymerase and Screening of Crystallization Conditions |
JING Jia-mei1,2,XUN Xin2,WANG Min1,PENG Ru-chao1,SHI Yi1,2,**( ) |
1 Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
2 Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China |
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Abstract The Lassa virus and the Machupo virus belong to the family of the Arenaviridae. They can cause human infection with high mortality and pose a threat to the public health. The arenavirus possesses a cap-snatching mechanism for transcription of viral genome, which is similar to that of other segmented negative-strand RNA viruses. This process involves cleavage of host mRNAs by an endonuclease (EN) domain located in the N-terminal region of the viral polymerase, and the cap-binding by the C-terminal region which has not been verified in Mammarenavirus. To this end, the recombinant expression vectors for the C-terminal region of Lassa virus and Machupo virus polymerase were constructed. The recombinant proteins were expressed in E. coli and was found that the proteins can exist with different oligomerization forms in solution.The oligomerization features by negative staining electron microscopy also were characterized. Finally, initial crystals of the C-terminal of Lassa virus polymerase were obtained, which provides a foundation for further study of its three-dimensional structure and functional mechanism.
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Received: 16 April 2019
Published: 15 January 2020
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Corresponding Authors:
Yi SHI
E-mail: shiyi@im.ac.cn
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