Herpes simplex virus type 1 ICP27 protein: its expression, purification and specific antiserum production.
Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni(2+)-NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.
Zhao
Lei
L
State Key Laboratory, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Ren
Xiao-ming
XM
Zheng
Alan C
AC
D000818:Animals; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D019085:Fluorescent Antibody Technique, Direct; D015870:Gene Expression; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D014709:Vero Cells
D004279:DNA, Viral; C057120:ICP27 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D011994:Recombinant Proteins