Eficiência na extração de DNA de diferentes tecidos de carpas através do protocolo de cloreto de sódio (NaCl)

With advances of farming and research in the area of molecular genetics, studies that can help with obtaining good quality DNA are necessary. The objective was to evaluate the efficiency of DNA extraction in different types of tissues of three species of carp by using the protocol of DNA extraction with Sodium Chloride (NaCl). The samples were subjected to DNA extraction, and the integrity was visualized on 1.5% agarose gel. In a total of 72 samples used for DNA extraction, all of them were positive, confirmed by the presence of bands on agarose gel. The capacity of amplifying the extracted DNA was tested by amplification reactions using the RAPD (random amplification of polymorphic DNA) technique, confirming that the DNA was of good quality for use in subsequent studies with molecular markers.Com o avanço da piscicultura e das pesquisas na área da genética molecular, existe a necessidade de estudos que possam aperfeiçoar a obtenção de DNA com boa qualidade. Diante disso, o objetivo desse trabalho foi avaliar a eficiência da extração de DNA em diferentes tipos de tecidos de três espécies de carpas, através do protocolo de extração de DNA com cloreto de sódio (NaCl). As amostras foram submetidas à extração de DNA e a integridade foi visualizada em gel de agarose 1,5%. Em um total de 72 amostras utilizadas na extração de DNA, todas foram positivas, confirmadas com a presença de banda no gel de agarose. A capacidade de amplificação do DNA extraído foi testada através de reações de amplificação utilizando a técnica RAPD (polimorfismos de DNA amplificados ao acaso), confirmando que o DNA apresenta boas condições para utilização em estudos posteriores com marcadores moleculares

Study of the variability of mhc genes b-l ß chicken (Gallus gallus domesticus) birds in brazilian caipira

The Brazilian caipira chickens egg blue are the result of random mating between different chicken breeds found in Brazil with chickens Araucania, giving rise to Brazilian caipira chicken with characteristic blue eggs. It stands out for its hardiness, disease resistance. And d adverse weather conditions and diet. The disease resistance is related to genetic variability of immune system genes. The immune response of chickens to pathogens is triggered from the presentation of antigens of the effect or cells of the immune system. This action is made by antigen-present cells (B cells, macrophages and a small percentage of T cells) animals that have surface molecules (glycoprotein) in wich these antigens are complexes. These glycoproteins presenting antigens, molecules called Major Histocompatibility Comples (MHC or B complex) can be class I or II. The class II, which present exogenous antigens, are composed of two polypeptide chains, an α chain and a ß chain. The chain is monomorphic α chain and ß is very polymorphic and is considered responsible for encoding different MHC class II molecules inn chickens. This study aimed to investigate the genetic variability of two gene loci of the MHC class II, B-LßI and B-LßII in caipiras chickens Brazilian blue eggs by obtaining sequences of these two genes and thus detect B-Lß existing alleles for these genes in these birds. This work was initially developed with DNA samples from 100 caipiras chickens blue eggs that were amplified from the region of interest and analyzed. From this analysis, the animals were grouped according to the sequence of exon2. Then, 16 animals were (with at least one representative from each group) for the reaction of DNA amplification, cloning and sequencing. The sample were submitted to the initiators of the genes B-LßII. Of these 16 birds were obtained 15 different sequences that were aligned to the standard sequence the B-LB12major (AJ248676) for BLßII and B-LB12c (AJ248578) to B-LßI, being eleven sequences B-LßII and B-LßI four sequences. Of these eleven sequences of the gene B-LßII seven were already described in the literature and four sequences of the gene B-LßII seven were already described in the literature and four sequences are new is not described, the other four sequences of the gene B-LßI three had already been described in the literature and a new . The 15 nucleotide sequences were transformed into sequences of amino acids. In the amino acid sequences of the gene was observed that B-LßI remains much more conserved than gene B-LßII. Thus this paper presents the genetic variability at both the DNA and polypeptide chain, genes B-LßII caipira chickens in Brazil s blue eggsAs galinhas caipiras brasileiras de ovos azuis são o resultado de cruzamentos aleatórios entre diversas raças de galinhas encontradas no Brasil com galinhas Araucanas, dando origem a uma galinha caipira brasileira com característica de ovos azuis. Destaca-se pela sua rusticidade, resistência a doenças e as condições adversas de clima e alimentação. A resistência a doenças esta relacionada à variabilidade genética dos genes do sistema imune. A resposta imune das galinhas a patógenos é desencadeada a partir da apresentação de antígenos as células efetoras do sistema imune. Esta ação é feita pelas células apresentadoras de antígenos (células B, macrófagos e uma pequena porcentagem de células T) dos animais que possuem moléculas de superfície (glicoproteínas) na qual são complexados estes antígenos. Estas glicoproteínas apresentadoras de antígenos, chamadas de moléculas do Complexo Maior de Histocompatibilidade (MHC ou Complexo B), podem ser de classe I ou de classe II. As de classe II, que apresentam antígenos exógenos, são formadas por duas cadeias polipeptídicas, uma cadeia α e uma cadeia β. A cadeia α é monomorfica e a cadeia β é bem polimórfica, e é considerada a responsável por codificar diferentes moléculas MHC classe II em galinhas. Este estudo teve como objetivo investigar a variabilidade genética em dois loci gênicos do MHC de classe II, B-LI e B-LII, em galinhas caipiras brasileiras de ovos azuis através da obtenção das sequências destes dois genes B-L e assim detectar os alelos existentes para estes genes nestas aves. Este trabalho foi desenvolvido inicialmente com amostras de DNA de 100 galinhas caipiras de ovos azuis que foram amplificadas para a região de interesse e analisadas. A partir desta análise, os animais foram agrupados de acordo com a sequência do exon 2. Depois, 16 animais foram selecionados (com pelo menos um representante de cada grupo) para a reação de amplificação do DNA, clonagem e sequenciamento. As amostras foram submetidas aos iniciadores dos genes B-LβI e B-LβII. Destas 16 aves obteve-se 15 diferentes sequências que foram alinhadas a sequência padrão B-LB12major (AJ248576) para o B-LβII e B-LB12c (AJ248578) para o B-LβI, sendo onze sequências B-LβII e quatro sequências B-LβI. Destas onze sequências do gene B-LβII sete já estavam descritas na literatura e quatro são sequências novas ainda não descritas, das outras quatro sequências do gene B-LβI três já haviam sido descritas na literatura e uma nova . As 15 sequências nucleotídicas encontradas foram deduzidas em sequências de aminoácidos. Nas sequências de aminoácidos foi observado que o gene B-LβI se mantém bem mais conservado do que o gene B-LβII. Assim, este trabalho apresenta a variabilidade genética dos genes B-LβI e BLβII tanto em nível de DNA quanto de cadeia polipeptídica, em galinhas caipiras brasileiras de ovos azui

Genetic polymorphism of fifteen microsatellite loci in Brazilian (blue-egg Caipira) chickens

The purpose of this study was to investigate the genetic polymorphism of fifteen microsatellites loci in Brazilian (blue-egg Caipira) chickens. Samples were collected from 100 blue eggs of Caipira chickens from rural properties in the city of Dois Lajeados, RS. After DNA extraction, the fragments related to molecular markers LEI0248, LEI0221, LEI0214, LEI0192, LEI0217, LEI0254, LEI0194, LEI0212, MCW0371, ADL0278, LEI0234, MCW0183, MCW0216, MCW0330 and MCW0081 were obtained by polymerase chain reaction (PCR). The statistical analysis were carried out with the softwares ARLEQUIN 3.5 version and CERVUS 3.0.3 version. The allelic and genotypic frequencies, deviations from Hardy-Weinberg equilibrium, estimates of observed (HO) and expected (HE) heterozygosity and polymorphic information content (PIC) were obtained for each marker locus. A total of 186 alleles from 15 loci were obtained, with sizes ranging of 83 to 490 base pairs. The medium number of alleles was 12.4, the HE was 0.76±0.14 and HO was 0.49±0.21 and PIC was 0.706. The first conclusion is that the microsatellites used are polymorphic and can be used to genetic studies in chickens. The second is that the "Caipira" chicken (blue eggs) population investigated has a great genic variability, which makes than an important source of genetic resources for future animal breeding programs

A global analysis of CNVs in swine using whole genome sequence data and association analysis with fatty acid composition and growth traits

Copy number variations (CNVs) are important genetic variants complementary to SNPs, and can be considered as biomarkers for some economically important traits in domestic animals. In the present study, a genomic analysis of porcine CNVs based on next-generation sequencing data was carried out to identify CNVs segregating in an Iberian x Landrace backcross population and study their association with fatty acid composition and growthrelated traits. A total of 1,279 CNVs, including duplications and deletions, were detected, ranging from 106 to 235 CNVs across samples, with an average of 183 CNVs per sample. Moreover, we detected 540 CNV regions (CNVRs) containing 245 genes. Functional annotation suggested that these genes possess a great variety of molecular functions and may play a role in production traits in commercial breeds. Some of the identified CNVRs contained relevant functional genes (e.g., CLCA4, CYP4X1, GPAT2, MOGAT2, PLA2G2A and PRKG1, among others). The variation in copy number of four of them (CLCA4, GPAT2, MOGAT2 and PRKG1) was validated in 150 BC1_LD (25% Iberian and 75% Landrace) animals by qPCR. Additionally, their contribution regarding backfat and intramuscular fatty acid composition and growth–related traits was analyzed. Statistically significant associations were obtained for CNVR112 (GPAT2) for the C18:2(n-6)/C18:3(n-3) ratio in backfat and carcass length, among others. Notably, GPATs are enzymes that catalyze the first step in the biosynthesis of both triglycerides and glycerophospholipids, suggesting that this CNVR may contribute to genetic variation in fatty acid composition and growth traits. These findings provide useful genomic information to facilitate the further identification of trait-related CNVRs affecting economically important traits in pigs.info:eu-repo/semantics/publishedVersio

Genotypic characterization of ten microsatellite loci in two Brazilian free range (Caipira) chicken lines

This study aimed to investigate the genetic variability of two Brazilian free range (Caipira) chickens lines using microsatellites analysis of ten loci. It was collected a total of 99 blood samples, which 49 were from Paraiso Pedres (PP) and 50 were from Rubro Negra (RN) lines. The amplification of the DNA fragments was performed by polymerase chain reaction (PCR) and the genotyping was conduct using ABI 3130 sequencer. The allele number variation was among 3 (LEI0254) to 32 (LEI0212) in the PP line, and 4 (LEI0254) to 31 (LEI0212) in the RN line. The allelic average per locus was 13.3 and 13.1 in the PP and RN lines, respectively. The average observed and the expected heterozygosity were 0.650 and 0.820 in the PP line, and 0.671 and 0.804 in the RN line. All of the analyzed loci were informative (PIC>0.5). These results indicate that these free-range animals have a high genetic variability, at least for the majority of the analyzed loci, and this genetic variation is higher than the commercial chickens and similar for the no-commercial birds

A global analysis of CNVs in swine using whole genome sequence data and association analysis with fatty acid composition and growth traits

<div><p>Copy number variations (CNVs) are important genetic variants complementary to SNPs, and can be considered as biomarkers for some economically important traits in domestic animals. In the present study, a genomic analysis of porcine CNVs based on next-generation sequencing data was carried out to identify CNVs segregating in an Iberian x Landrace backcross population and study their association with fatty acid composition and growth-related traits. A total of 1,279 CNVs, including duplications and deletions, were detected, ranging from 106 to 235 CNVs across samples, with an average of 183 CNVs per sample. Moreover, we detected 540 CNV regions (CNVRs) containing 245 genes. Functional annotation suggested that these genes possess a great variety of molecular functions and may play a role in production traits in commercial breeds. Some of the identified CNVRs contained relevant functional genes (e.g., <i>CLCA4</i>, <i>CYP4X1</i>, <i>GPAT2</i>, <i>MOGAT2</i>, <i>PLA2G2A</i> and <i>PRKG1</i>, among others). The variation in copy number of four of them (<i>CLCA4</i>, <i>GPAT2</i>, <i>MOGAT2</i> and <i>PRKG1</i>) was validated in 150 BC1_LD (25% Iberian and 75% Landrace) animals by qPCR. Additionally, their contribution regarding backfat and intramuscular fatty acid composition and growth–related traits was analyzed. Statistically significant associations were obtained for CNVR112 (<i>GPAT2</i>) for the C18:2(n-6)/C18:3(n-3) ratio in backfat and carcass length, among others. Notably, GPATs are enzymes that catalyze the first step in the biosynthesis of both triglycerides and glycerophospholipids, suggesting that this CNVR may contribute to genetic variation in fatty acid composition and growth traits. These findings provide useful genomic information to facilitate the further identification of trait-related CNVRs affecting economically important traits in pigs.</p></div