Peptide sequences obtained from cyanogen bromide fragments of bovine lactoperoxidase (bLPO) were used to design oligonucleotide probes for library screening. These probes were used to screen a cDNA library constructed from bovine mammary tissue. Three overlapping clones were obtained, the longest of which (T3) contained a reading frame of 712 amino acid residues. The encoded amino acid sequence was homologous to those recently reported for myelo-, thyro-, and eosinophil peroxidases. Two possible amino termini of the mature enzyme were identified, and the predicted mature protein matched previous molecular weight estimates of 78,500. Of eight bovine tissues tested, transcription of T3 sequences were detected in mammary tissue only. Using the bLPO cDNA as a probe, a single hybridizing clone was found in a human mammary gland cDNA library. This clone (M1) encoded the carboxy-terminal 324 residues of a peroxidase distinct from the other three known human peroxidases, and was closely related to bLPO. This result confirms the presence of at least one distinct lactoperoxidase in humans.
DNA amplification reactions using long, non-random, single oligonucleotide primers with thermostable DNA polymerase and avian genomic DNA generate unique, reproducible multiband patterns of DNA fragments. These DNA fragments are produced from amplification reactions using single primers of at least twentyfive bases in length which were derived from the chicken α A -globin gene. The patterns of amplified DNA fragments were polymorphic among individual chickens. Within families, polymorphic band patterns were found to be heritable, as the DNA fragments of progeny were present in at least one of the parents. In a conventional polymerase chain reaction containing a pair of primers, the relative concentrations of the primers can be manipulated to produce either a discrete two primer product or a multiband pattern identical to the single primer products. In addition, the number of bands in the multiband pattern increases as the stringency of the annealing step of the DNA amplification is decreased. The products are believed to result from recognition of a family of related sequences in the genome. This single primer amplification method has general application in the establishment of molecular relatedness, discovery of new genetic markers, resolution of questions of paternity, identification of individual organisms, and measurement of biodiversity within and among populations.
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