While genotyping wild red deer (Cervus elaphus) at microsatellite loci for paternity assignment, we found three loci (MAF65, BOVIRBP and CelJP23) with segregating nonamplifying alleles. Nonamplifying alleles were detected through mismatches between known mother-offspring pairs and by significant deviations from Hardy-Weinberg equilibria. In a wide range of molecular ecology application, and especially in parentage assignment, the possible existence of undetectable alleles must be taken into account; this may be particularly important for microsatellite data.

We have developed a technique to establish catalogues of protein products of arrayed cDNA clones identified by DNA hybridisation or sequencing. A human fetal brain cDNA library was directionally cloned in a bacterial vector that allows IPTG-inducible expression of His6-tagged fusion proteins. Using robot technology, the library was arrayed in microtitre plates and gridded onto high-density in situ filters. A monoclonal antibody recognising the N-terminal RGSH6sequence of expressed proteins (RGS.His antibody, Qiagen) detected 20% of the library as putative expression clones. Two example genes, GAPDH and HSP90alpha, were identified on high-density filters using DNA probes and antibodies against their proteins.

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Molecular techniques have enabled behavioural ecologists to reassess mating systems from a genetic perspective. Studies of paternity frequently reveal that mating behaviour does not always reflect parentage, and may bring to light alternative mating tactics. Here we present a comparison of behavioural and genetic measures of male reproductive success in a mammalian mating system in which both sexes are highly promiscuous. Rather than having a stable harem social structure, Soay rams (Ovis aries) on the island of Hirta, St Kilda, UK usually consort with individual oestrous ewes sequentially. Not all matings occur between consort pairs, however, and ewes have been seen to mate with up to 10 different rams on the same day. Using locus-specific polymorphism at five protein and 10 microsatellite DNA loci, we determined paternity for 236 lambs born into three cohorts, and compared paternity with estimates of mating success derived from more than one census of rutting behaviour. The correlation between the number of ewes with which each ram was witnessed in consort and the number of paternities assigned was positive and statistically significant, and rams that were observed in consort with a ewe were 18 times more likely to have sired her offspring than other candidate rams. However, most lambs (73%) were not sired by a ram seen in consort with the oestrous mother. Many juveniles, yearlings and some adult rams were rarely seen in consort with ewes, yet were assigned a significant number of paternities. These results suggest that mating tactics differ between age groups, and that alternative mating strategies among adults that do not involve forming consorts with many females also confer mating success. For these reasons, census-based observations of consort associations between individuals cannot be used to accurately estimate individual male reproductive success in this population.

We investigated polymorphism at protein and microsatellite DNA loci in an isolated, umnanaged and cyclic population of Soay sheep on the island of Hirta, St. Kilda. Extensive molecular variation was revealed at both protein loci (mean heterozygosity 7.78 per cent) and microsatellite loci (mean heterozygosity 50.93 per cent). Typically, large mammals possess limited protein variation and we were surprised to observe such a level of protein heterozygosity, particularly considering the genetic history of the Hirta population. Indeed, compared to other mammals, Soay sheep lie within the top 17 per cent of the distribution of average protein heterozygosities. We discuss the level of protein heterozygosity in the context of other mammalian species, other breeds of sheep and the genetic history of Soay sheep. Possible explanations for a large average and interlocus variance in protein heterozygosity are proposed. Although little data are available from other studies to compare with microsatellite DNA variability in this population, we discuss the potential application of microsatellite markers to interpopulation and interspecific genetic studies.