A SspI PCR-RFLP detecting a silent allele at the goat CSN2 locus

The comparison between the cDNA sequence obtained and the published sequences of the goat CSN2 alleles showed a new single nucleotide polymorphism (SNP) (transition C-T) at the 180th nucleotide of the ninth exon. This mutation, which took place at 124 nt from the polyadenylation site, identifies a silent allele at the CSN2 locus named CSN2 A1. Since the 9th exon C-T transition creates a SspI endonuclease restriction site, the SspI digestion of a PCR product of 360 bp spanning the 9th exon and flanking regions, would allow carriers for the presence of thymine to be identified. The allelic frequency of the CSN2 A1 allele, determined in 170 goats belonging to an undefined genetic type reared in the province of Naples (Italy), was 0.23 It has been observed that the sequences in the 3’ untranslated regions (UTR), proximal to the polyadenylation site, can affect the mechanism of mRNA deadenylation and degradation. Therefore, it is reasonable to hypothesize that the C-T transition might, directly or indirectly, influence the stability of the mRNA and, consequently, the amount of protein produced

Genetic polymorphism of goat CSN1S1 and CSN1S2 genes and their impact on milk composition

by means of selection based on parent genotyping at the DNA level it will be possible to obtain goat populations producing milk characterised by the absence of either as1-casein, or s2-casein or -casein. Such milk could be useful for specific technological processes of transformation or for specific nutritional or dietary purposes in order to attenuate the negative consequence of some metabolic deficiencies and allergies and to contribute to the prevention of some diseases Considering the remarkable quantitative polymorphism characterizing the main protein fractions encoding gene, some goat milk types are more similar to the woman milk and, therefore, could be used, with better results, in the human feeding. In fact, the woman milk possess particular characteristics. It is lacking in the -lactoglobulin fraction and it is characterized by a general low content in casein, in particular, as2 casein absence and as1 casein traces, similar to the milk produced by goats with genotype CSN1S2 0/0

A point mutation in the splice donor site of intron 7 in the as2-casein encoding gene of the Mediterranean River buffalo results in an allele-specific exon skipping

The CSN1S2 cDNA of 10 unrelated Mediterranean River Buffaloes reared in Southern Italy was amplified by RT-PCR, while the region from the 6th to the 8th exon of the CSN1S2 gene was amplified from genomic template. cDNA sequence comparisons showed that five individuals had a normal transcript only (named CSN1S2A), one had a deleted transcript only (named CSN1S2B), because of the splicing out of the 27-bp of exon 7, and the remaining four had a heterozygous pattern. Analysis of the genomic sequences revealed a FM865620: g.773G>C transversion that caused inactivation of the intron 7 splice donor site and, consequently, the allele-specific exon skipping characteristic of the CSN1S2B allele. The g.773G>C mutation creates a new AluI restriction site enabling a PCR– RFLP rapid genotyping assay. The cDNA sequences showed three additional exonic mutations forming an extended haplotype with the g.773G>C polymorphism: FM865618: c.459C>T, c.484A>T and c.568A>G homozygous and heterozygous respectively in the CSN1S2BB and CSN1S2AB buffaloes. The first is silent, while the remaining two are non-conservative (p.Ile162Phe and p.Thp200Ala respectively). The genotype frequencies (37 CSN1S2A/A, 15 CSN1S2A/B and one CSN1S2B/B) are in agreement with Hardy–Weinberg equilibrium, with the frequency of the deleted B allele being 0.16. The predicted bubaline as2B protein is 198 aa long instead of 207 aa and would also be characterized by the presence of Phe at position 147 and Ala at 185

Direito societário e do mercado de capitais em debate: barragens, crise ambiental e responsabilidade

- Divulgação dos SUMÁRIOS das obras recentemente incorporadas ao acervo da Biblioteca Ministro Oscar Saraiva do STJ. Em respeito à Lei de Direitos Autorais, não disponibilizamos a obra na íntegra.- Localização na estante: 347.725:504(81) D598s- Organizado por: Pedro A. L. Ramunno

Contencioso societário: e novas perspectivas do direito empresarial

- Divulgação dos SUMÁRIOS das obras recentemente incorporadas ao acervo da Biblioteca Ministro Oscar Saraiva do STJ. Em respeito à Lei de Direitos Autorais, não disponibilizamos a obra na íntegra.- Localização na estante: 347.7(81) C761s- Organizado por: Pedro A. L. Ramunno

Genotyping at the CSN1S1 locus by PCR-RFLP and AS-PCR in a Neapolitan Goat Population

The goat CSN1S1 gene has for many years been an excellent model for demonstrating that most of the variability observed in the as1-casein content in goat’s milk is due to the presence of autosomal alleles at a single structural locus. Until now, about 17 alleles associated to at least four levels of as1-casein expression in milk have been described at the CSN1S1 locus in the domestic goat (Capra hircus). The great importance of goat as1-casein polymorphism is due to its qualitative as well as quantitative implications. In the present work five PCR protocols (PCR-RFLPs, AS-PCR) were set up for rapid genotyping of B1, B2*, B3, B4 and C CSN1S1 alleles, until nowdetectable only by milk electrophoresis. Application of these protocols, together with previously described methods to identify CSN1S1 01, E, M, F, N and A* (CSN1S1 A, G, I, H) alleles, allow us to define, at DNA level, the genetic structure of the autochthonous goat reared in the province of Naples for the highest number of possible alleles at this locus. Monitoring of CSN1S1 variability in the Neapolitan goat population indicates a high frequency of low (F, 0.368) and null (N, 0.227) alleles