Genomic analysis of human multigene families using chromosome-specific vectorette PCR.
We report a technique for the rapid determination of genomic structure of individual members of human interspersed multigene families which circumvents the requirement for genomic clone isolation. In this approach, vectorette libraries were constructed from human/rodent somatic cell hybrid DNA harbouring single members of the gene family. Using these libraries as PCR templates with nested gene-specific primers in combination with a common vectorette primer resulted in the amplification of gene-specific products suitable for the subsequent determination of intron/exon structure. We have applied this technique to characterise members of two gene families.
Moynihan
T P
TP
Molecular Medicine Unit, University of Leeds, St James's University Hospital, UK.
Markham
A F
AF
Robinson
P A
PA
D002875:Chromosomes / Q000235:genetics; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells; D008025:Ligases / Q000032:analysis / Q000737:chemistry; D005810:Multigene Family / Q000235:genetics*; D016133:Polymerase Chain Reaction; D044763:Ubiquitin-Conjugating Enzymes
D017931:DNA Primers; D044763:Ubiquitin-Conjugating Enzymes; D008025:Ligases