Expression and purification of thioredoxin-his6-ZmDREB2.7 fusion protein in Escherichia coli for raising antibodies
Thuy Linh Nguyen1, 2, Thuy Duong Nguyen1, Van Hai Nong1, Thi Thu Hue Huynh1*
1Institute of Genome Research, Vietnam Academy of Science and Technology
2University of Science, Vietnam National University, Hanoi
Received 3 August 2018; accepted 23 November 2018
Dehydration-responsive element-binding (DREB) proteins play a critical role in the plant’s drought-tolerance mechanism despite their presence in minor amounts in the cell. In this study, a maize-derived transcription factor protein, ZmDREB2.7, was overexpressed in the Escherichia coli strain Rosetta 1. The interested gene conjugating with the thioredoxin gene (TrxA) and his6 tag in the pET-32a vector encoded a 55.7 kDa fusion protein. The optimum condition for inducing the thioredoxin-his6-ZmDREB2.7 expression was five hours of induction with 0.05mM IPTG at 30°C. The Tris-HCl 20 mM pH 8.0 lysis buffer was harnessed to extract the recombinant protein for the purification process. Using the immobilized-metal affinity chromatography column, the recombinant protein was purified and then injected into rabbits. The antisera containing polyclonal antibodies (pAbs) could specifically recognize the ZmDREB2.7 fusion protein. This study represents updated data on the bacterial expression of the recombinant ZmDREB2.7 protein and the production of anti-ZmDREB2.7 pAbs.
Keywords: E. coli, fusion expression, recombinant protein, ZmDREB2.7 protein.
Classification numbers: 3.1