Purification of Recombinant Tagged Proteins

The protein expression plasmids with the target cloned in pET30a(+), pMAL-c5X-His, and pGEX-4T3 were transformed into BL21/DE3 pLysS E.coli bacteria (#L1195, Promega, Madison, WI). The recombinant His-, MBP-, Strep-, and GST-tagged proteins were purified as previously described [23 (link),49 (link),87 (link),88 (link)] or purified with Bio-Rad NGC Chromatography System (Bio-Rad Laboratories) using Bio-Scale Mini Nuvia IMAC Cartridges (#7800811, Bio-Rad, CA). The His-tagged NS1-70 was further purified by running through a HiLoad Superdex 200 pg column (#28989335, Cytiva) with Bio-Rad NGC Chromatography System at a flow rate of 0.5 ml/min as previously described [49 (link)].

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Ning K., Wang Z., Cheng F., Yan Z, & Qiu J. (2022). The small nonstructural protein NP1 of human bocavirus 1 directly interacts with Ku70 and RPA70 and facilitates viral DNA replication. PLoS Pathogens, 18(6), e1010578.

Publication 2022

Bio-Rad NGC Chromatography System Bio-Scale Mini Nuvia IMAC Cartridges HiLoad Superdex 200 pg column

Bacteria Chromatography E coli Imac Plasmids Promega Protein target Proteins Strep

Corresponding Organization : University of Kansas Medical Center

Other organizations : University of Iowa

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Variable analysis

independent variables

Protein expression plasmids with the target cloned in pET30a(+), pMAL-c5X-His, and pGEX-4T3

dependent variables

Purification of recombinant His-, MBP-, Strep-, and GST-tagged proteins

control variables

Transformation of the plasmids into BL21/DE3 pLysS E. coli bacteria

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