The Chinese hamster HPRT gene: Restriction map, sequence analysis, and multiplex PCR deletion screen

摘要: Abstract The fine structure of the Chinese hamster hypoxanthine guanine phosphoribosyltransferase (HPRT) gene has been determined; nine exons and is dispersed over 36 kb DNA. Exons 2–9 are contained within overlapping λ bacteriophage clones exon 1 was obtained by an inverse polymerase chain reaction (PCR). All have sequenced, together with their immediate flanking regions, these sequences compared to those mouse human HPRT genes. Sequences immediately all but first show considerable homology between different species region around less conserved, apart from preserved location putative functional elements. Oligonucleotide primers derived were used amplify simultaneously seven exon-containing fragments in a multiplex PCR. This simple procedure identify total partial deletions among HPRT-deficient mutants. PCR quicker perform than Southern analysis, traditionally study such mutants, also provides specific for further analysis. often as target mutation studies vitro because ease selection forward reverse mutants; information presented here will enhance means investigating molecular defects this gene.