Cloning and Prokaryotic Expression of Pathogenesis-related protein 1 Gene from Oriental Hybrid Lily Sorbonne

doi:10.7522/j.issn.1000-694X.2017.00031

Journal of Desert Research ›› 2018, Vol. 38 ›› Issue (3): 584-591.DOI: 10.7522/j.issn.1000-694X.2017.00031

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Cloning and Prokaryotic Expression of Pathogenesis-related protein 1 Gene from Oriental Hybrid Lily Sorbonne

Wang Le^1,2, Yang Liu^1,2, Guo Zhihong¹, Zhang Yubao¹, Wang Yajun¹, Yang Guo¹, Xie Zhongkui¹

1. Gaolan Station of the Agricultural and Ecological Experiment, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2016-12-29 Revised:2017-04-01 Online:2018-05-20 Published:2018-11-06

Abstract

Abstract: Pathogenesis-related (PR) proteins are widely involved in plant defense against both biotic and abiotic stresses. In the present study, we characterized a PR1 protein from the oriental hyrid lily cultivar Sorbonne via rapid amplification of cDNA ends (RACE), and designated as LhSorPR1. The cDNA full-length of LhSorPR1 isolated was 870 bp, consisting a 5' untranslate region (UTR) of 47 bp and a 3' UTR of 274 bp. The predicted open reading frame (ORF) of LhSorPR1 was 549 bp, encoded a protein of 182 amino acids. The mature LhSorPR1 (devoid of a signal peptide) had a theoretical isoelectric point of 5.69 and a calculated molecular weight of 18.19 kDa. The three dimentional (3D) model of LhSorPR1 was bulit by homology modelling, and the bulit model showed high similarity to its template p14a. The similarity in 3D strucutures indicated the conservation of protein function of PR1 proteins in plants. The pCold Ⅱ-LhSorPR1 recombinant vector was constructed to express LhSorPR1 in E.coli, and the His₆-tagged recombinant protein was expressed as inclusion bodies in the insoluble fraction under IPTG induction. The His₆-labeled recombinant LhSorPR1 protein was verified by immunoblot, and purified by chromatography.

Key words: lily, disease resistance, pathogenesis-related protein, homology modelling, prokaryotic expression

CLC Number:

Q943

Wang Le, Yang Liu, Guo Zhihong, Zhang Yubao, Wang Yajun, Yang Guo, Xie Zhongkui. Cloning and Prokaryotic Expression of Pathogenesis-related protein 1 Gene from Oriental Hybrid Lily Sorbonne[J]. Journal of Desert Research, 2018, 38(3): 584-591.

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Cloning and Prokaryotic Expression of Pathogenesis-related protein 1 Gene from Oriental Hybrid Lily Sorbonne

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