High-level expression, one-step purification of soluble Ad5-knob protein and its activity assay.
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.
Yang
Juyun
J
Xiangya Hospital of Central South University, Changsha, China.
Zhao
Yanzhong
Y
Liu
Ting
T
Chen
Yuxiang
Y
Yu
Shuyi
S
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
D008670:Metals; D011994:Recombinant Proteins; D004247:DNA