Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

doi:10.3969/j.issn.1000-2006.2016.03.007

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2016, Vol. 40 ›› Issue (03): 41-46.doi: 10.3969/j.issn.1000-2006.2016.03.007

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Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

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

The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Lin'an 311300, China

Online:2016-06-18 Published:2016-06-18

Abstract

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.

CLC Number:

Q518.2

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, 2016, 40(03): 41-46.

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Expression and purification of Phyllostachys violascens SOC1 protein in prokaryotic system

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