PMID: 7523540Jun 1, 1994Paper

Distinction of mouse interferon-alpha subtypes by polymerase chain reaction utilizing consensus primers and type-specific oligonucleotide probes

Journal of Interferon Research
T K HughesP L Rady

Abstract

We have developed a novel method to study the subtype-specific expression of interferon-alpha (IFN-alpha) in the mouse system: we synthesized and used consensus oligonucleotide primers to allow simultaneous polymerase chain reaction (PCR) amplification of multiple murine IFN-alpha gene sequences. In addition, a set of subtype-specific oligomer probes were designed and used to distinguish between IFN-alpha genes that differ by only a few bases. The consensus primers, corresponding to two regions highly conserved among murine IFN-alpha subtypes, were used in reverse transcription and PCR amplification of total cellular RNA, isolated from IFN-gamma-treated murine L-929 cells, to yield a fragment of the anticipated approximately 520-bp size. Southern analysis of the amplified product using an internal consensus oligomer probe confirmed specific amplification of murine IFN-alpha gene(s). Subtype-specific oligonucleotide probes indicate that IFNs-alpha 1, -alpha 2, and -alpha 5 are present following IFN-gamma treatment, whereas IFN-alpha 4 remains virtually absent. Our results indicate that the expression of specific IFN-alpha subtypes may be subject to complex regulation, dependent upon inducing agents and cell types involved, and c...Continue Reading

Citations

Aug 28, 2004·Brain, Behavior, and Immunity·John HunzekerJohn F Sheridan
Aug 30, 2003·Brain, Behavior, and Immunity·Griselle C OrtizPhillip T Marucha
Oct 4, 2005·Arthritis and Rheumatism·Jianwei LiChandra Mohan
Aug 3, 1999·Biological Chemistry·M MatsumotoT Taniguchi
Jul 27, 2007·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Gong ChengZhaoxin Zheng
Jan 1, 1996·Biotherapy·R M FriedmanS Baron

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