title: Prokaryotic expression of pathogenesis related protein 1 gene from Nicotiana benthamiana: antifungal activity and preparation of its polyclonal antibody.
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.
authors.affiliation: Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, Sichuan University, Chengdu, 610064, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhu
authors.forename: Moyun
authors.initials: M
authors.lastname: Xu
authors.forename: Shaodong
authors.initials: S
authors.lastname: Wang
authors.forename: Shudan
authors.initials: S
authors.lastname: Jia
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Honghui
authors.initials: H
authors.lastname: Lin
authors.forename: Dehui
authors.initials: D
authors.lastname: Xi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification; D000935:Antifungal Agents / Q000276:immunology / Q000494:pharmacology*; D001688:Biological Products / Q000276:immunology / Q000494:pharmacology*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D014026:Nicotiana / Q000235:genetics*
chemical_list: D000906:Antibodies; D000935:Antifungal Agents; D001688:Biological Products; D018744:DNA, Plant; D010940:Plant Proteins; D011994:Recombinant Proteins; C053376:pathogenesis-related proteins, plant