A 11.7-kb deletion triggers intersexuality and polledness in goats.

Mammalian sex determination is governed by the presence of the sex determining region Y gene (SRY) on the Y chromosome. Familial cases of SRY-negative XX sex reversal are rare in humans, often hampering the discovery of new sex-determining genes. The mouse model is also insufficient to correctly apprehend the sex-determination cascade, as the human pathway is much more sensitive to gene dosage. Other species might therefore be considered in this respect. In goats, the polled intersex syndrome (PIS) mutation associates polledness and intersexuality. The sex reversal affects exclusively the XX individuals in a recessive manner, whereas the absence of horns is dominant in both sexes. The syndrome is caused by an autosomal gene located at chromosome band 1q43 (ref. 9), shown to be homologous to human chromosome band 3q23 (ref. 10). Through a positional cloning approach, we demonstrate that the mutation underlying PIS is the deletion of a critical 11.7-kb DNA element containing mainly repetitive sequences. This deletion affects the transcription of at least two genes: PISRT1, encoding a 1.5-kb mRNA devoid of open reading frame (ORF), and FOXL2, recently shown to be responsible for blepharophimosis ptosis epicanthus inversus syndrome (BPES) in humans. These two genes are located 20 and 200 kb telomeric from the deletion, respectively.

Analysis of genetic relationships between 10 cattle breeds with 17 microsatellites.

To guide genetic conservation programmes with objective criteria, general genetic variability has to be taken into account. This study was conducted to determine the genetic variation between 10 cattle breeds by using 17 microsatellite loci and 13 biochemical markers (11 blood groups, the transferrin and beta-casein loci). Microsatellite loci were amplified in 31-50 unrelated individuals from 10 cattle breeds: Charolais, Limousin, Breton Black Pied, Parthenais, Montbéliard, Vosgien, Maine-Anjou, Normande, Jersey and Holstein. Neighbor-joining trees were calculated from genetic distance estimates. The robustness of tree topology was obtained by bootstrap resampling of loci. A total of 210 alleles of the 17 microsatellites were detected in this study and average heterozygosities ranged from 0.53 in the Jersey breed to 0.66 in the Parthenais breed. In general, low bootstrap values were obtained: with the 17 microsatellites, the highest bootstrap values concerned the Holstein/Maine-Anjou grouping with an occurrence of 74%; with the biochemical markers, this node had an occurrence of 79% and the Charolais/Limousin grouping appeared with an occurrence of 74%; when microsatellites and biochemical polymorphism were analysed together, the occurrence of the Holstein/Maine-Anjou grouping was 90% and that of the Charolais/Limousin grouping was 42%. These results suggest that 30 microsatellites, a number currently considered as sufficient to distinguish closely related breeds is, in fact, probably insufficient.

A microsatellite within the bovine kappa-casein gene reveals a polymorphism correlating strongly with polymorphisms previously described at the protein as well as the DNA level.

The polymorphism of a (TA)n(CA)n repeat microsatellite present in the third intron of the bovine kappa-casein gene (CASK) has been investigated. The existence of six alleles differing only in the number of dinucleotide repeats has been established. A total of 330 animals belonging to nine different pure bred Bos taurus French breeds or to a cross-bred Bos taurus x Bos indicus population (Créole) were genotyped. The distribution of the microsatellite alleles was examined and clear breed differences were noted. Genotyping of animals by isoelectric focusing (IEF) or restriction fragment length polymorphism (RFLP) (TaqI) was performed, in order to examine the relationship of the microsatellite polymorphism to other previously described CASK polymorphisms, at the protein and DNA levels. Strong correlation was seen, indicating that evolution of the various polymorphisms was not independent, and nine CASK haplotypes were observed.

Two of the three genetic variants of goat alpha s1-casein which are synthesized at a reduced level have an internal deletion possibly due to altered RNA splicing.

This paper describes the elucidation of the primary structure of the three genetic variants of goat alpha s1-casein, alpha s1-Cn D, E and F, which have been found to be associated with reduced amounts of alpha s1-casein in milk. Variant E has the same electrophoretic mobility as variant B, but differs from the latter by the substitutions of Arg for Lys and of Thr for Ala at positions 100 and 195. A genetically controlled event which does not affect the amino acid sequence of this variant might be responsible for its lower rate of synthesis compared to that of alpha s1-casein B. The deletion of 11 amino acids at positions 59-69 and of 37 amino acids at positions 59-95 in variant B leads to variants D and F. In both cases the deletions, which start at the same position of the polypeptide chain, include the major phosphorylation site of the protein. On the basis of sequence data for casein genes and cDNAs, it was concluded that the deletions occurring in the D and F variants are due to the exclusion of one and several exons, respectively. The observed deletions in the proteins could thus be the consequence of splice site mutations which would induce altered RNA processing and hence reduce the rate of synthesis of the casein.

Sequence of caprine alpha s1-casein and characterization of those of its genetic variants which are synthesized at a high level, alpha s1-CnA, B and C.

The sequence of caprine alpha s1-casein (199 residues) was established. The peptide chain has the same length as, and shows a 88% degree of identity with, its bovine counterpart. With the ovine alpha s1-casein, the sequence of which was deduced from that of its mRNA, the degree of identity is 97%, counting as one difference a deletion of eight residues in the ovine protein. The differences between the three genetic variants associated with a high alpha s1-casein content in milk are simple substitutions. Variant alpha s1-CnA differs from variant alpha s1-CnB by two substitutions, 16 Leu (A)----Pro (B) and 77 Gln (A)----Glu (B), the latter inducing the appearance of a phosphate group on 75 Ser. Variant alpha s1-CnC differs from alpha s1-CnB by three substitutions, 8 His (B)----Ile (C), 100 Arg (B)----Lys (C) and 195 Thr (B)----Ala (C). The original type of caprine alpha s1-casein could be another hypothetical genetic variant, having the same electrophoretic mobility as alpha s1-CnB.

The C system of cattle blood groups. 2. Partial genetic map of the system.

20 cases of irregular inheritance of phenogroups in the C system of cattle blood groups were used to deduce a partial genetic map of this system, taking into consideration the 11 internationally recognized antigenic factors and the 4 additional factors recently described by Grosclaude et al. (1980). This partial map bears resemblance to that established by Bouw et al. (1974) in Dutch cattle. The operational length of the DNA sequence coding for the C system was estimated to be 0.3 centimorgan, a value which is approximately half of that obtained for the B system by Grosclaude et al. (1979). It is concluded that the phenogroups of the C system. like those of the B system, are controlled by a cluster of loci.

The C system of cattle blood groups. 1. Additional factors in the system.

Four additional cattle blood group antigenic factors, provisionally termed F1, F6, F10 and F15, were shown to belong to the C system. Factor F1 appears to be a linear subtype of C" (initially designated F2, or P1B1). It is suggested that future international nomenclature should adopt C"1 and C"2 in place of F1 and C". No phenogroup was found to include C" together with C2 or C1, but a few phenogroups lack the three factors. Thus C1, C2 and C" do not form a closed system within the C system as concluded by Duniec et al. (1973). The effectiveness of the additional factors to uncover the genetic variability of the C system, and to translate phenotypes into genotypes is exemplified in the Charolais breed.

A genetic and biochemical analysis of a polymorphism of bovine alpha S2-casein.

Using gel electrophoresis a genetic polymorphism of alpha S2-casein (Cn) was discovered in individual milk samples from 2 bovine breeds of the eastern part of France (Vosgienne and Montbéliarde). The 3 observed phenotypes (Plate 1) are determined by 2 co-dominant alleles at an autosomal locus. The alpha S2-Cn A variant was the only one known up to now in European breeds (reference variant) and alpha S2-Cn D is a new variant, whose bands overlap the beta-casein A band at pH 8.6, and migrate faster than alpha S2-Cn A at pH 3.0. The sequence of the polypeptide chain alpha S2-Cn D differs from that of alpha S2-Cn A by the deletion of a very acidic nonapeptide, which includes a cluster of 3 phosphoseryl residues. Due to the characteristics of the reference sequence, this deletion cannot be exactly located but it involves residues 50-58, or 51-59, or 52-60. A genetic analysis shows that locus alpha S2-Cn is closely linked to the cluster alpha S1-Cn--beta-Cn--kappa-Cn. The 4 casein species are thus synthesized by 4 closely linked loci.

The genetic map of the B system of cattle blood groups as observed in French breeds.

40 cases of irregular inheritance of phenogroups in the B system of cattle blood groups, that were presumed to have arisen from single crossing-over, have been used to establish a partial genetic map of this system. In addition, the relative positions in the map of several antigenic factors were inferred from relationships observed between particular phenogroups occurring in French breeds. The tentative map thus obtained shows an overall similarity to that established by Ruiterkamp et al. (1977) in Dutch Friesian cattle. This result, in addition to other arguments, supports the hypothesis that the genetic structure of the B system of cattle blood groups is basically the same in all taurine breeds. Evidence is given, for the first time, for the occurrence in the B system of genetic events other than single crossing-over (double crossing-over or gene conversion, and possibly deletion). The rate of recombination between the genetic determinants of the terminal factors of the system, Q and I', was calculated in the progeny of some Normande bulls heterozygous for these two determinants (447 gametes); a value of 1.34 centimorgan was obtained. However complementary data indicate that 0.7 centimorgan would be a better estimate.

Analysis of alloantisera against bovine lymphocytes. Joint report of the 1st International Bovine Lymphocyte Antigen (BoLA) workshop.

The results and agreements of the 1 international BoLA workshop, held in Edinburgh, Scotland in August 1978, are reported. Most of these concern the results from a comparison test of 249 alloantisera to bovine lymphocytes, the antisera being contributed by 9 laboratories. These sera were compared directly in Edinburgh on a panel of lymphocytes from 130 cattle of 21 breeds. In the microlymphocytotoxicity test used 75% of the sera reacted. Sixty eight of these sera were grouped into clusters according to their reaction patterns against the lymphocyte panel. Eleven of these clusters were clearly defined and were given workshop BoLA designations. In addition 22 sera were assigned to subgroups of the agreed clusters. There was no evidence that the method of production of the sera had any effect on their specificity. Although genetic data was not available, the phenotypes of the test panel of lymphocytes are consistent with the clusters detecting antigens controlled by multiple alleles at a single autosomal locus. It was agreed to name the genetic region where this putative locus is located BoLA (bovine lymphocyte antigen).