Cloning and expression of the nucleocapsid protein gene of nipah virus in different strains of Escherichia coli.
The coding region of the nucleocapsid (N) gene was amplified from the viral RNA and inserted into the bacterial expression vector, pTrcHis2, for intracellular expression in three Escherichia coli strains: TOP 10, BL 21 and SG 935. The N protein was expressed as a fusion protein containing the myc epitope and His-tag at its C-terminal end. The amount of the fusion protein expressed in strain SG 935 was significantly higher than the other two strains, and was detected by the anti-myc antibody, anti-His and swine anti-NiV serum. Hence, the N(fus) protein produced in E. coli could serve as an alternative antigen for the detection of anti-NiV in swine.
Ong
Swee Tin
ST
Department of Biochemistry and Microbiology, Faculty of Science and Environmental Studies, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
Tan
Wen Siang
WS
Hassan
Sharifah Syed
SS
Mohd Lila
Mohd Azmi
MA
Yusoff
Khatijah
K
D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000302:isolation & purification; D018098:Paramyxovirinae / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000235:genetics
D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins; D015678:Viral Structural Proteins