Intragenic haplotypes at the bovine CSN1S1 locus

A new alternative genotyping method based on PCR-SSCP was developed for direct differentiation of the CSN1S1 alleles B and C in the coding region. In addition a PCR-RFLP test based on a MaeIII restriction site in the promoter region of CSN1S1, reported in the literature as an alternative test for the differentiation of CSN1S1*B and C was used and the alleles named b and c. Genotyping of 649 animals belonging to 17 European and Turkish cattle breeds showed differences in occurrence and frequency of the alleles. CSN1S1*B occurred in all breeds with frequencies varying from 0.50 in Anatolian Blackup to 1.0 in e.g. Ayrshire. CSN1S1*b on the other hand varied from 0.63 in Jersey, 0.97 in Ayrshire to 1.0 in e.g. Angler. Comparison of the results from both typing methods and positions in the gene showed that both mutations do not always occur together. From the resulting four intragenic haplotypes (B-b, B-c, C-c and C-b) B-b is predominant in all breeds with frequencies varying from 0.3450 in Anatolian Black to 1.0 in Angler and Scottish Highland. The number of haplotypes varied from only one in Angler and Scottish Highland, two in Ayrshire, three in Asturian Valley and Turkish Grey Steppe to all four in the other 12 breeds. Correlation between allele frequencies and the geographic origin of the breeds was significant for the MaeIII promoter polymorphism.O. Jann, E.-M. Prinzenberg, H. Brandt, J. L. Williams, P. Ajmone-Marsan, P. Zaragoza, C. Özbeyaz, and G. Erhard

Genetic variation in the kappa-casein gene (CSN3) of Chinese yak (Bos grunniens) and phylogenetic analysis of CSN3 sequences in the genus Bos

Variants of kappa-casein (CSN3) have been extensively studied in cattle and 13 alleles have been identified at the protein and DNA levels to date. Evolution of some of these alleles and a possible common ancestor remain unclear. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of CSN3 exon IV in domesticated yak revealed a 2-allele polymorphism showing migration patterns different from known cattle variants. The PCR products of both yak CSN3 SSCP alleles were sequenced. All yak had nucleotide sequences coding for Thr in AA position 136 (identical to bovine CSN3*A) and Ala in position 148 (identical to bovine CSN3*B). The sequencing results were confirmed by PCR-RFLP analysis using HindIII and TaqI. A 12-bp insertion in the coding region, representing a repeated nucleotide and AA motif, was found in 1 yak allele. The duplication corresponds to the codons for AA 147 to 150 (Glu-Ala-Ser-Pro) or 148 to 151 (Ala-Ser-Pro-Glu), which are repeated identically. In 21 yak samples genotyped by PCR-SSCP analysis, frequencies for the insertion variant and the short variant were about 68 and 32%, respectively. The loss of the insertion may have led to the ancestral CSN3 allele from which all currently known variants of CSN3 in the genus Bos evolved. This is the first report of polymorphisms in the yak CSN3 gene and may be helpful for future studies on genetic variation within and between yak populations or on associated traits

Caprine \u3ba-Casein (CSN3) polymorphism : new developments in molecular knowledge

A high degree of polymorphism was recently found at the kappa-casein (CSN3) locus in the domesticated goat (Capra hircus). In the present study, 2 new patterns previously identified by PCR-single-strand conformation polymorphism analysis (SSCP) were characterized. The allele provisionally named "X" (GenBank Accession no. AY350425) differs from CSN3*C (AF485341) by a (silent) A -> G substitution at position 509 of the goat CSN3 reference sequence (X60763). As this newly identified sequence changes the amino acid sequence, and the already known CSN3*C allele (AF485341) has an additional silent mutation, we proposed a change in nomenclature to reflect these changes, indicating the silent mutation with the prime symbol (i.e., '). The CSN3*M allele (provisionally named "Y") results in a new protein variant, differing by 2 nonsynonymous mutations from the CSN3*F allele. The new variant is characterized by a G -> A transition at nucleotide position 384, resulting in the amino acid exchange Asp(90)-> Asn(90), and a C -> T transition at position 550, resulting in a Val(145)-> Ala(145) substitution. Thus, the number of alleles identified in the domesticated goat has increased to 16, of which 13 are protein variants and 3 are silent mutations, involving a total of 15 polymorphic sites in CSN3 exon 4. Data on the distribution of the main alleles in 7 goat breeds of Europe, West Africa, and the Near East show differences in the occurrence and frequency of the alleles between breeds and geographic origin with the highest number of alleles found in goat breeds from the Near East

Caprine κ-casein (CSN3) polymorphism: new developments of the molecular knowledge

A high degree of polymorphism was recently found at the κ-casein (CSN3) locus in the domesticated goat (Capra hircus). In the present study, 2 new patterns previously identified by PCR-single-strand conformation polymorphism analysis (SSCP) were characterized. The allele provisionally named “X” (GenBank Accession no. AY350425) differs from CSN3*C (AF485341) by a (silent) A→G substitution at position 509 of the goat CSN3 reference sequence (X60763). As this newly identified sequence changes the amino acid sequence, and the already known CSN3*C allele (AF485341) has an additional silent mutation, we proposed a change in nomenclature to reflect these changes, indicating the silent mutation with the prime symbol (i.e., ′). The CSN3*M allele (provisionally named “Y”) results in a new protein variant, differing by 2 nonsynonymous mutations from the CSN3*F allele. The new variant is characterized by a G→A transition at nucleotide position 384, resulting in the amino acid exchange Asp90→ Asn90, and a C→T transition at position 550, resulting in a Val145→Ala145 substitution. Thus, the number of alleles identified in the domesticated goat has increased to 16, of which 13 are protein variants and 3 are silent mutations, involving a total of 15 polymorphic sites in CSN3 exon 4. Data on the distribution of the main alleles in 7 goat breeds of Europe, West Africa, and the Near East show differences in the occurrence and frequency of the alleles between breeds and geographic origin with the highest number of alleles found in goat breeds from the Near East

Marker-assisted conservation of European cattle breeds: an evaluation

Two methods have been developed for the assessment of conservation priorities on the basis of molecular markers. According to the Weitzman approach, contributions to genetic diversity are derived from genetic distances between populations. Alternatively, diversity within and across populations is optimized by minimizing marker-estimated kinships. We have applied, for the first time, both methods to a comprehensive data set of 69 European cattle breeds, including all cosmopolitan breeds and several local breeds, for which genotypes of 30 microsatellite markers in 25-50 animals per breed have been obtained. Both methods were used to calculate the gain in diversity if a breed was added to a set of nine non-endangered breeds. Weitzman-derived diversities were confounded by genetic drift in isolated populations, which dominates the genetic distances but does not necessarily increase the conservation value of a breed. Marker-estimated kinships across populations were less disturbed by genetic drift than the Weitzman diversities and assigned high conservation values to Mediterranean breeds, which indeed have genetic histories that differ from the non-endangered breeds. Prospects and limitations of marker-assisted decisions on conservation priorities are discussed