Cloning and functional characterization of a eucaryotic DNA photolyase gene from Neurospora crassa.
We cloned a genomic fragment of a photolyase gene from Neurospora crassa by polymerase chain reaction using synthesized oligonucleotide primers designed from the most conserved amino acid sequences among photolyases of various organisms. Using the cloned fragment as a hybridization probe we isolated a genomic fragment and cDNA clones encoding the complete photolyase gene of this organism. The amino acid sequence of the photolyase deduced from the determined nucleotide sequence indicates a protein consisting of 615 amino acid residues (Mr 69,971), which is most similar to that of Saccharomyces cerevisiae. Like yeast photolyase it contains a protruding amino terminus which is missing in photolyases of bacterial origin. Comparison of amino acids sequences among six photolyases suggests that the Neurospora crassa photolyase is more similar to photolyases of pterin type than those of deazaflavin type.
Yajima
H
H
Research Institute for Tuberculosis and Cancer, Tohoku University, Sendai, Japan.
Inoue
H
H
Oikawa
A
A
Yasui
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015342:DNA Probes / Q000235:genetics; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology; D005800:Genes, Fungal / Q000235:genetics*; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D014466:Ultraviolet Rays
D015342:DNA Probes; D009841:Oligonucleotides; D004255:Deoxyribodipyrimidine Photo-Lyase