Abstract
The nucleotide sequence of the pathogenesis-related protein 1(PR-1) gene was obtained from Nicotiana benthamiana using RT-PCR. Restriction enzyme cutting sites of EcoRI and NotI were introduced to the ORF fragments of PR-1, they were then linked together with pET-30a (+) and transformed into E. coli BL21 (DE3). The target protein was induced by 1.5 mM IPTG, at 37°C for 4 h. The expressed protein was purified by Ni–NTA and an anti-NbPR-1 polyclonal antibody was prepared using rabbits. The antibody could detect the expression of PR-1 in N. benthamiana and other Nicotiana plants. NbPR-1 protein has four α-helices and two β-sheets by homology modeling. Furthermore, the purified NbPR-1 protein exhibited a broad-spectrum antifungal activity.
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Acknowledgments
This work was supported by the National Nature Science Foundation of China (91017004, 30970214, 31070210, 31071669, and 31171835), National Key Basic Research ‘973’ Program of China (2009CB118500), Fundamental Research Funds for the Central Universities (2011SCU04B34) and Doctoral Foundation of the Ministry of Education (20110181110059 and 20070610172).
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Zhu, F., Xu, M., Wang, S. et al. Prokaryotic expression of pathogenesis related protein 1 gene from Nicotiana benthamiana: antifungal activity and preparation of its polyclonal antibody. Biotechnol Lett 34, 919–924 (2012). https://doi.org/10.1007/s10529-012-0851-5
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DOI: https://doi.org/10.1007/s10529-012-0851-5