title: High-level expression, one-step purification of soluble Ad5-knob protein and its activity assay.
abstract: This report describes a simple, highly efficient, and reproducible method for obtaining large quantities of highly pure recombinant Ad5-knob protein, which can be used for gene-delivery application. The Ad5- knob protein expressed in Escherichia coli contained a His tag at the N-terminus that allowed one-step isolation by immobilized metal affinity chromatography (IMAC). The activity of the recombinant protein was tested by receptor-binding assay in Hela cells for potential application in gene delivery.
authors.affiliation: Xiangya Hospital of Central South University, Changsha, China.
authors.forename: Juyun
authors.initials: J
authors.lastname: Yang
authors.forename: Yanzhong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Ting
authors.initials: T
authors.lastname: Liu
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Chen
authors.forename: Shuyi
authors.initials: S
authors.lastname: Yu
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002845:Chromatography; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D015316:Genetic Therapy / Q000295:instrumentation* / Q000379:methods*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008670:Metals / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D008670:Metals; D011994:Recombinant Proteins; D004247:DNA