Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

doi:10.3969/j.issn.1000-2006.2016.03.007

Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

TANG Yegen,SHI Quan, LIN Xinchun, XU Yingwu

Journal of Nanjing Forestry University (Natural Sciences Edition） ›› 2016, Vol. 40 ›› Issue (03) : 41-46.

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Journal of Nanjing Forestry University (Natural Sciences Edition） ›› 2016, Vol. 40 ›› Issue (03) : 41-46. DOI: 10.3969/j.issn.1000-2006.2016.03.007

Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

TANG Yegen,SHI Quan, LIN Xinchun, XU Yingwu^*

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Abstract

In order to obtain soluble recombinant PvSOC1(Phyllostachys violascens SUPPERSSOR OF CO OVEREXPRESSION 1)protein in vitro. We used pET-GST as expression vector, to over-express full-length and IKC structural domain of PvSOC1 in E.coli Rosetta prokaryotic system. We found the optimal expression strategy is when the suspension culture's OD₆₀₀ value reaches 0.4-0.6, add IPTG and keep it's concentration at 0.4 mmol/L. Kept incubating for 5 h at the temperature of 37℃ to induce the protein.We found IKC domain is soluble,to compare with the fullength sequence,it started from No.62 amino acid and ends at No.225. Added TEV(tobacco etch virus protease)protease to remove GST tag.Combined glutathione sepharose resin with size exclusion chromatography(SEC)to get highly purified IKC domain of PvSOC1.And chromatography data proof this domain exists as a octamer.

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TANG Yegen,SHI Quan, LIN Xinchun, XU Yingwu. Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system[J]. Journal of Nanjing Forestry University (Natural Sciences Edition）. 2016, 40(03): 41-46 https://doi.org/10.3969/j.issn.1000-2006.2016.03.007

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