Isolation of cDNAs encoding the catalytic domain of poly(ADP-ribose) polymerase from Xenopus laevis and cherry salmon using heterologous oligonucleotide consensus sequences.
We have isolated and sequenced cDNAs encoding the catalytic domain of poly(ADP-ribose) polymerase (PARP) from Xenopus laevis and Oncorhyncus masou (cherry salmon). The cDNAs were amplified by polymerase chain reaction using heterologous oligonucleotides corresponding to the conserved sequences of mammalian cDNAs as primers. The deduced amino acid sequences of Xenopus laevis and cherry salmon cDNA showed 84.4% and 75.6% similarities to that of human PARP, respectively. In both species, mRNA for PARP was identified as a single band of 4 kb, and PARP mRNA was abundant in ovary and brain. Thus, mixed oligonucleotide-primed amplification is a useful method in the cloning of cDNAs from different species, and the catalytic domain of PARP is conserved structurally among phylogenetically different species, suggesting an importance of poly(ADP-ribosyl)ation.
Department of Biochemistry, University of Tsukuba, Japan.
Y
Y
Ozawa
K
K
Uchida
M
M
Uchida
Y
Y
Ami
S
S
Kushida
N
N
Okada
M
M
Miwa
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D016384:Consensus Sequence; D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012474:Salmon; D017386:Sequence Homology, Amino Acid; D014982:Xenopus laevis
D009838:Oligodeoxyribonucleotides; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases