Generation of polymerase chain reaction-specific probes for library screening using single degenerate primers.
Degenerate oligonucleotide primers were made to peptide sequences from hydroxylamine oxidoreductase (HAO) from Nitrosomonas europaea. The primers were used singly in PCR reactions to amplify portions of the gene for HAO from genomic DNA. Southern hybridizations using fragments amplified with each primer showed that they labeled the same genomic DNA fragments. The PCR-amplified fragments were successfully used to screen a gene library for clones containing the HAO gene. The method of isolating genes by PCR with single primers has general utility.
Hommes
N G
NG
Laboratory for Nitrogen Fixation Research, Oregon State University, Corvallis 97331-2902, USA.
Arp
D J
DJ
Sayavedra-Soto
L A
LA
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D015723:Gene Library; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009606:Nitrosomonas / Q000201:enzymology / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D010088:Oxidoreductases; C021401:hydroxylamine dehydrogenase