title: Quantitative and qualitative analysis of human IgG subclass specific mRNA using solution hybridization.
abstract: Four IgG subclasses have been identified in the human system. Despite the fact that they exhibit differences in their functional properties, antigenic properties and chemical composition and demonstrate age-related shifts in their expression, the genes coding for their constant regions show extensive homology at the nucleotide level (greater than 95%). The only parts of the C gamma genes that show significant variation among the different IgG subclasses are the exons coding for their hinge regions (less than 60%). Taking advantage of such sequence variation, we have developed specific RNA probes which allowed us to analyse the expression of the C gamma 1, C gamma 2, C gamma 3 and C gamma 4 genes at the mRNA level using solution hybridization. We have defined optimal conditions that allow the detection of picogram levels of IgG subclass specific mRNA, while keeping the cross-reactivity between the different probes below 2%. This approach will significantly facilitate studies aiming at characterizing the molecular mechanisms regulating the expression of the four human IgG heavy chain constant region genes.
authors.affiliation: Unit for Applied Cell and Molecular Biology, Umeå University, Sweden.
authors.forename: P
authors.initials: P
authors.lastname: Sideras
authors.forename: L
authors.initials: L
authors.lastname: Nilsson
authors.forename: K B
authors.initials: KB
authors.lastname: Islam
authors.forename: H
authors.initials: H
authors.lastname: Ericsson
authors.forename: L
authors.initials: L
authors.lastname: Hammarström
authors.forename: C I
authors.initials: CI
authors.lastname: Smith
mesh_terms: D001483:Base Sequence; D006801:Humans; D007074:Immunoglobulin G / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*
chemical_list: D007074:Immunoglobulin G; D012333:RNA, Messenger