Characterization of the chicken and rat DCoH gene domains using an improved ligation-mediated PCR method.
The bifunctional protein DCoH (dimerization cofactor of HNF-1) is a transcriptional coactivator of homeodomain proteins and a cytoplasmic enzyme. Here we report on the isolation, nucleotide sequence and genomic structure of the chicken and rat DCoH genes. They have an unusual first exon coding for a single amino acid. The promoters are located within CpG-islands and lack TATA and CAAT consensus sequences. Analysis of the number of CpG dinucleotides in the promoter indicates little changes since man and chicken were separated in evolution whereas a 67% reduction was observed in the rat lineage. The DNA sequences were obtained entirely without cloning. For walking into unknown genomic regions we describe a modification of the ligation-mediated polymerase chain reaction (LM-PCR) that can replace conventional screening and cloning methods and greatly expedite the isolation of genomic DNA.
Bossow
S
S
Adolf-Butenandt-Institut für Physiologische Chemie, Physikalische Biochemie und Zellbiologie der Universität München, Germany.
Riepl
S
S
Igo-Kemenes
T
T
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D018076:DNA, Complementary; D006836:Hydro-Lyases / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D006836:Hydro-Lyases; C076392:pterin-4a-carbinolamine dehydratase