Solubility, immunogenicity and physical properties of the nucleocapsid protein of Nipah virus produced in Escherichia coli.
The nucleocapsid (N) protein of Nipah virus (NiV) can be produced in three Escherichia coli strains [TOP10, BL21(DE3) and SG935] under the control of trc promoter. However, most of the product existed in the form of insoluble inclusion bodies. There was no improvement in the solubility of the product when this protein was placed under the control of T7 promoter. However, the solubility of the N protein was significantly improved by lowering the growth temperature of E. coli BL21(DE3) cell cultures. Solubility analysis of N- and C-terminally deleted mutants revealed that the full-length N protein has the highest solubility. The soluble N protein could be purified efficiently by sucrose gradient centrifugation and nickel affinity chromatography. Electron microscopic analysis of the purified product revealed that the N protein assembled into herringbone-like particles of different lengths. The C-terminal end of the N protein contains the major antigenic region when probed with antisera from humans and pigs infected naturally.
Tan
Wen Siang
WS
Department of Biochemistry and Microbiology, Faculty of Science and Environmental Studies, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. wstan@fsas.upm.edu.my
Ong
Swee Tin
ST
Eshaghi
Majid
M
Foo
Sze-Shir
SS
Yusoff
Khatijah
K
D000818:Animals; D000956:Antigens, Viral; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D045464:Henipavirus Infections / Q000276:immunology / Q000821:virology; D006801:Humans; D008854:Microscopy, Electron; D045405:Nipah Virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017384:Sequence Deletion; D012995:Solubility
D000956:Antigens, Viral; D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins