Coxsackie Virus B3 Gene Cloning

Coxsackie virus B3 genes (Nancy strain, GenBank: M33854.1) were individually PCR-amplified from a genomic plasmid using primers containing restrictions site overhangs, digested, and ligated into the pcDNA4/TO plasmid (Invitrogen) that has been engineered to fuse a 2xStrep tag to the C-terminus of proteins cloned into the multiple cloning site^{47 (link)}. Mutations for 2A C107A and 3C C147A were introduced by site-directed mutagenesis.

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Diep J., Ooi Y.S., Wilkinson A.W., Peters C.E., Foy E., Johnson J.R., Zengel J., Ding S., Weng K.F., Laufman O., Jang G., Xu J., Young T., Verschueren E., Kobluk K.J., Elias J.E., Sarnow P., Greenberg H.B., Hüttenhain R., Nagamine C.M., Andino R., Krogan N.J., Gozani O, & Carette J.E. (2019). Enterovirus Pathogenesis Requires the Host Methyltransferase SETD3. Nature microbiology, 4(12), 2523-2537.

Publication 2019

pcDNA4/TO plasmid

A proteins Genomic Mutations Plasmid Primers Proteins c Site directed mutagenesis Strain Virus genes

Corresponding Organization :

Other organizations : Stanford University, University of California, San Francisco, Chan Zuckerberg Initiative (United States), Quantitative BioSciences

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Variable analysis

independent variables

Mutations for 2A C107A and 3C C147A

dependent variables

Protein expression with 2xStrep tag

control variables

PcDNA4/TO plasmid (Invitrogen) that has been engineered to fuse a 2xStrep tag to the C-terminus of proteins cloned into the multiple cloning site

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