Detection of differential expression of mouse interferon-alpha subtypes by polymerase chain reaction using specific primers.
Specific primers for nine mouse interferon-alpha (IFN-alpha) subtypes, namely, IFN-alpha1, IFN-alpha1-9, IFN-alpha2, IFN-alpha4, IFN-alpha5, IFN-alpha7, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB, were designed and evaluated on Poly(I).Poly(C)-induced and influenza virus-infected L929 cells. Specificity of the primers was confirmed in a cross-polymerase chain reaction (cross-PCR). IFN-alpha1, IFN-alpha1-9, IFN-alpha4, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB were found to be induced in L929 cells 6-9 h after Poly(I).Poly(C) treatment. The amplification of a particular subtype was not biased in the presence of excess of other templates. Differential expression of the IFN-alpha subtypes was observed in influenza A/NWS/33- and B/Lee/40-infected L929 cells. A/NWS/33 virus was found to upregulate the gene expression of IFN-alpha1, IFN-alpha4, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB in L929 cells as early as 6 h after infection. In B/Lee/40-infected L929 cells, only IFN-alpha4 was upregulated. Our results suggest that the designed primers will serve as a useful tool in analyzing the expression of IFN-alpha subtypes in various systems and hence for the evaluation of their function.
Fung
M C
MC
Department of Biology, The Chinese University of Hong Kong, Hong Kong, China.
Sia
S F
SF
Leung
K N
KN
Mak
N K
NK
D000818:Animals; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics; D005347:Fibroblasts; D009980:Influenza A virus / Q000276:immunology; D009981:Influenza B virus / Q000276:immunology; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D016898:Interferon-alpha; D012333:RNA, Messenger