Mapping of human H chain V region genes (VH4) using deletional analysis and pulsed field gel electrophoresis

Abstract

We have used molecular genetic mapping techniques to establish the order and approximate chromosomal locations of VH4 elements on both chromosomes 14 from a single patient. A total of 10 BglII restriction fragments carrying VH4 elements was characterized. The genomic order of VH4-carrying restriction fragments was determined by analysis of the pattern of loss of hybridizing bands from the genomes of a panel of monoclonal lymphoblastoid cell lines which had well characterized rearrangements of the Ig locus on each chromosome. Some individual elements were identified using sequence-specific oligonucleotide probes. Physical dimensions were estimated by the assignment of these ordered elements to large (50-350-kb) restriction fragments using two-dimensional pulse field gel electrophoresis. Six such fragments spanning approximately 890 kb were physically linked and ordered. The chromosomes differed with respect to the complement of VH4 elements, although no evidence was found of major differences in organization. The establishment of a panel of well characterized deletion lines facilitates the rapid mapping of defined restriction fragments carrying VH elements.