Rapid detection of immunoglobulin gene somatic hypermutation using heteroduplex formation.
In order to identify somatic hypermutation of rearranged human immunoglobulin genes, we have examined heteroduplex formation between cloned VH6 genes. In test systems, the presence of five or more point mutations could be detected by examining the formation of heteroduplexes between a known germline VH6 gene and other sequenced genes using polyacrylamide gel electrophoresis. If a mutated sequence was used, then the presence of two or more mutations could be detected. The method was used for rapid screening of VH6-D-JH rearrangements for the presence of point mutations before sequencing, and to distinguish between different highly mutated sequences, allowing clones containing the same rearrangement to be identified indirectly.
Department of Clinical Immunology, Flinders University of South Australia, Bedford Park.
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Nicholson
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Zola
D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics*; D005803:Genes, Immunoglobulin / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes; D017354:Point Mutation
D009692:Nucleic Acid Heteroduplexes