title: The recombinant expression and activity detection of MAF-1 fusion protein.
abstract: This study establishes the recombinant expression system of MAF-1 (Musca domestica antifungal peptide-1) and demonstrates the antifungal activity of the expression product and shows the relationship between biological activity and structure. The gene segments on mature peptide part of MAF-1 were cloned, based on the primers designed according to the cDNA sequence of MAF-1. We constructed the recombinant prokaryotic expression plasmid using prokaryotic expression vector (pET-28a(+)) and converted it to the competent cell of BL21(DE3) to gain recombinant MAF-1 fusion protein with His tag sequence through purifying affinity chromatographic column of Ni-NTA. To conduct the Western Blotting test, recombinant MAF-1 fusion protein was used to produce the polyclonal antibody of rat. The antifungal activity of the expression product was detected using Candida albicans (ATCC10231) as the indicator. The MAF-1 recombinant fusion protein was purified to exhibit obvious antifungal activity, which lays the foundation for the further study of MAF-1 biological activity, the relationship between structure and function, as well as control of gene expression.
authors.affiliation: Department of Parasitology, Pathogen Biology Laboratory, Guizhou Medical University, No 9 Beijing Road, Guiyang, Guizhou Province, 550004, P.R.China.
authors.forename: Ping
authors.initials: P
authors.lastname: Fu
authors.affiliation: Department of Parasitology, Pathogen Biology Laboratory, Guizhou Medical University, No 9 Beijing Road, Guiyang, Guizhou Province, 550004, P.R.China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Internal Medicine, Center for Cardiovascular Biology and Atherosclerosis Research, The University of Texas Medical School at Houston, 6431 Fannin Street, MSB 1.246; Houston, Texas, 77030, USA.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Parasitology, Pathogen Biology Laboratory, Guizhou Medical University, No 9 Beijing Road, Guiyang, Guizhou Province, 550004, P.R.China.
authors.forename: Guo
authors.initials: G
authors.lastname: Guo
authors.affiliation: Department of Parasitology, Pathogen Biology Laboratory, Guizhou Medical University, No 9 Beijing Road, Guiyang, Guizhou Province, 550004, P.R.China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Parasitology, Pathogen Biology Laboratory, Guizhou Medical University, No 9 Beijing Road, Guiyang, Guizhou Province, 550004, P.R.China.
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006793:Houseflies; D051258:MafB Transcription Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000935:Antifungal Agents; D051258:MafB Transcription Factor; C490405:Mafb protein, rat; D015513:Oncogene Proteins; D011993:Recombinant Fusion Proteins