RESUMO
The polymorphism of 23 microsatellites in the four main cattle breeds in Belgium (Holstein Friesian, Belgian Blue, Belgian Red Pied and East Flemish) was analysed. Heterozygosity, polymorphism information content, the effective number of alleles, exclusion probability and the probability of genotypic identity for two random individuals were calculated for all microsatellites and all breeds. The Belgian Blue breed is generally a little less polymorphic in comparison with the other three breeds. Estimates of the genetic distances between these breeds confirmed the widely accepted proposition that the Belgian Blue is the most genetically distinct of these breeds. The three other breeds are likely to become one population, given current breeding strategies. Exclusion probabilities in parentage control cases are > 0.9999 in all four breeds when all 23 microsatellites are used and > 0.98 with only the two most polymorphic multiplexes.
Assuntos
Bovinos/genética , Variação Genética , Repetições de Microssatélites , Polimorfismo Genético , Animais , Marcadores Genéticos , Genótipo , Heterozigoto , Reação em Cadeia da Polimerase , Especificidade da EspécieAssuntos
Antígenos de Bactérias , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli , Proteínas de Fímbrias , Imunidade Inata/imunologia , Doenças dos Suínos/microbiologia , Animais , Antígenos de Superfície/imunologia , Aderência Bacteriana , Microvilosidades/microbiologia , SuínosRESUMO
A detailed physical map of the porcine MHC class III region on Chr 7 was constructed with a panel of probes in a series of hybridizations on genomic pulsed field gel electrophoresis (PFGE) Southern blots. A precise organization of the 700-kb segment of DNA between G18 and BAT1 can now be proposed, with more than 30 genes mapped to it. Comparison of this region with homologous regions in human and mouse showed only minor differences. The biggest difference was observed in the CYP21/C4 locus with only one CYP21 gene and one C4 gene found, whereas in human and mouse these genes are duplicated. These results show the class III region is very well conserved between pig, human, and mouse, in contrast with the class I and class II regions, which seem more prone to rearrangements.
Assuntos
Complexo Principal de Histocompatibilidade/genética , Suínos/genética , Animais , Mapeamento Cromossômico , Complemento C4/genética , Sistema Enzimático do Citocromo P-450/genética , DNA Complementar/genética , Humanos , Camundongos , Dados de Sequência Molecular , RNA Helicases , RNA Nucleotidiltransferases/genética , Especificidade da EspécieRESUMO
The RD gene, named after the arginine (R) and aspartic acid (D) repeat in the central part of its protein, was initially mapped in the mouse H-2S subregion between C4 and BF. It was later mapped in the same position in the human MHC and here we show it is also conserved in the pig MHC class III region, close to the complement BF gene. A pig RD genomic clone was isolated from a lambda-phage library. Hybridizations on genomic DNA separated with pulsed field gel electrophoresis identified common 220 kb NruI, 130 kb EagI and 200 kb MluI bands for RD, BF and C2. The RD gene has also a 17 kb Kp nI and 11 kb SacI fragment in common with BF but not with C2. The close linkage of the RD and BF genes was further established by hybridization of BF to a genomic lambda-phage clone also containing the RD gene. This genomic RD clone overlaps with a lambda-phage clone previously isolated and containing the complete BF gene and the 3' part of C2. The distance between RD and BF is about 6 kb. The junction between the two complement genes BF and C2 was sequenced and the BF 5' promoter region, overlapping the 3' noncoding region of C2, was compared with that of the human BF promoter. The overall homology was about 80% and all but one identified promoter elements were found in the same position in both genes. The results obtained demonstrate the RD-BF-C2 organization is strongly conserved between human, mouse and pig. No polymorphisms were detected in either the RD gene or in the BF promoter region using polymerase chain reaction and restriction fragment polymorphism analysis.
Assuntos
Complexo Principal de Histocompatibilidade , Suínos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , Complemento C2/genética , Proteínas do Sistema Complemento/genética , Sequência Conservada , Primers do DNA/genética , Ligação Genética , Humanos , Camundongos , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
Three genomic clones were isolated from a size-selected pig DNA library by hybridization with a DNA-fingerprint probe. Analysis at the sequence level revealed that all three clones contain interrupted stretches of triplet repeats mainly composed of CAC and CAT triplets. Evaluation of the corresponding loci for polymorphism by Southern blot hybridization showed considerable length variation. For two loci the polymorphism was also demonstrated by polymerase chain reaction (PCR) amplification. The PiGMaP reference pedigree was typed for all three loci.
Assuntos
Genoma , Polimorfismo Genético , Sequências Repetitivas de Ácido Nucleico , Suínos/genética , Animais , Sequência de Bases , Clonagem Molecular , Impressões Digitais de DNA , Sondas de DNA , Elementos de DNA Transponíveis , Biblioteca Gênica , Dados de Sequência Molecular , Mutagênese Insercional , Reação em Cadeia da Polimerase/métodosRESUMO
Three related alpha-protease inhibitors, PI2 I, PI3 C and PI4 C2, of blood serum of the pig (Sus scrofa) were isolated. PI2 I inhibited both trypsin and chymotrypsin; PI3 C and PI4 C2 strongly inhibited chymotrypsin, but did not significantly inhibit trypsin. By using SDS-PAGE, the three proteins were found to be composed of single polypeptide chains, and molecular weights were 63,000 for PI2 I, 58,000 for PI3 C and 64,000 for PI4 C2. All three proteins were shown to be glycoproteins. In PI3 C, eight sialic acid residues were found, and in PI4 C2 (similarly as in PI2 F) 10-11 residues were found. Amino acid composition as well as N-terminal sequences of the three proteins were very similar, indicating close homology. Comparison of these partial amino acid sequences with the cDNA-deduced amino acid sequence of pig alpha-antichymotrypsin (AACT; Buchman, 1989, GenBank, Accession No. M29508) revealed great similarities, the sequence of PI2 I being virtually identical with the pig AACT. On the basis of all available results, PI2 is proposed to be pig AACT, an orthologue of human AACT.
Assuntos
Proteínas Sanguíneas/química , Quimotripsina/antagonistas & inibidores , Inibidores de Proteases/sangue , alfa 1-Antiquimotripsina/sangue , Sequência de Aminoácidos , Aminoácidos/análise , Animais , DNA Complementar/química , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Peso Molecular , Neuraminidase/farmacologia , Homologia de Sequência , Suínos , Inibidores da Tripsina/sangue , alfa 1-Antiquimotripsina/químicaRESUMO
The BAT1 gene has previously been identified about 30 kb upstream from the tumor necrosis factor (TNF) locus and close to a NF kappa b-related gene of the nuclear factor family in the major histocompatibility complex (MHC) of human, mouse, and pig. We now show that the BAT1 translation product is the homolog of the rat p47 nuclear protein, the WM6 Drosophila gene product, and probably also Ce08102 of Caenorhabditis elegans, all members of the DEAD protein family of ATP-dependent RNA helicases. This family has more than 40 members, including the eukaryotic translation initiation factor-4A (eIF-4A), the human nuclear protein p68, and the Drosophila oocyte polar granule component vasa. BAT1 spans about 10 kb, is split into 10 exons of varying length, and encodes a protein of 428 amino acids (approximately 48 kDa). Human and pig BAT1 cDNAs display 95.6% identity in the coding region and 80% identity in the 5' and 3' noncoding regions. Several repeat sequences of different types were identified in introns of the porcine BAT1 gene. Three different mRNAs, 4.1, 1.7, and 0.9 kb, respectively, were detected in all tissues analyzed upon hybridization with porcine BAT1 cDNA. Transfection and expression of human BAT1 cDNA after tagging with a heterologous antibody recognition epitope revealed a nuclear localization of the hybrid protein. An MspI RFLP was detected in an SLA class I typed family, confirming the localization of the BAT1 gene in the porcine MHC. BAT1 thus encodes a putative nuclear ATP-dependent RNA helicase and is likely to have an indispensable function.
Assuntos
Genes , Complexo Principal de Histocompatibilidade , Família Multigênica , RNA Nucleotidiltransferases/genética , Suínos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular Transformada , Chlorocebus aethiops , Cruzamentos Genéticos , Drosophila melanogaster/genética , Feminino , Masculino , Dados de Sequência Molecular , Filogenia , RNA Helicases , Ratos/genética , Sequências Repetitivas de Ácido Nucleico , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Telômero/genética , TransfecçãoAssuntos
Mapeamento Cromossômico , DNA Satélite/análise , Suínos/genética , Animais , Sequência de Bases , Clonagem Molecular , Sequência Consenso , Feminino , Biblioteca Genômica , Hibridização in Situ Fluorescente , Masculino , Dados de Sequência Molecular , Linhagem , Sequências Repetitivas de Ácido NucleicoRESUMO
Twenty-seven (CA)n and two (GA)n microsatellite clones were isolated out of a size-selected genomic pig library. These were sequenced and the number of uninterrupted dinucleotides was found to range from 12 to 26. Flanking primers were chosen for 11 dinucleotide repeats and optimal conditions for polymerase chain reaction (PCR) amplifications were established. Different microsatellite loci were amplified simultaneously by combining primer sets. Related and unrelated pigs were screened for length polymorphisms of the different microsatellite loci. The polymorphic information content (PIC) of these loci ranged between 0.62 and 0.83. Segregation studies in pig reference families established Mendelian inheritance. Locus S0022 was found to be X-linked.
Assuntos
DNA Satélite/química , Polimorfismo Genético , Suínos/genética , Alelos , Animais , Composição de Bases , Sequência de Bases , Clonagem Molecular , Frequência do Gene , Biblioteca Gênica , Dados de Sequência Molecular , Linhagem , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Sequências Repetitivas de Ácido Nucleico , Homologia de Sequência do Ácido NucleicoRESUMO
A genomic clone, SSBf1, containing the complement factor B (BF), a major histocompatibility class III antigen, has been isolated from a porcine genomic library. Partial sequencing and comparison with a human BF gene has identified seven exons coding for amino acids of Ba and Bb, the two subunits of BF. The protein sequence similarity with the human BF is on the average 87%. Southern blot analysis confirmed the existence of only one BF gene per haploid genome. Restriction fragment length polymorphism typing with Taq I showed that there are at least three different porcine BF-haplotypes.
Assuntos
Clonagem Molecular , Fator B do Complemento/genética , Suínos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Complemento C2/genética , Dados de Sequência Molecular , Polimorfismo de Fragmento de RestriçãoRESUMO
Five bovine paternal half-sib pedigrees for a total of 527 individuals were typed for six blood group systems: A, B, F/V, L, S, Z; for nine biochemical polymorphisms: ADA, MPI, PGM-3(slow), NP, Gc, Pi2, Tf, Ptf1 and Ptf2; and for restriction fragment length polymorphisms at five autosomal loci: Tg, GH, LDLr, BoLA-DQ and BoLA-DY. Two of the pedigrees were informative for segregation at the 'muscular hypertrophy' locus, and one was informative at the coat colour determining 'roan' locus. Linkage analysis was performed between all markers. Linkage was demonstrated between the S and F/V blood group systems (z = 3.11), adding one locus to the previously identified linkage group VII (LGVII) [Pi-2 and S], the most likely order being Pi2-S-F/V with maximum likelihood recombination rates of 0.208 and 0.211. Also shown to be linked were the blood group systems B and Z (z = 5.7, theta = 0.245). We confirmed the observation previously made by Andersson et al. (1988) of a high recombination rate between class II genes DQ and DY, suggesting either a larger physical distance between those genes than expected from comparative data, or the presence of a 'recombinational hotspot' in the bovine major histocompatibility complex. No linkage was found either with the 'muscular hypertrophy' locus, or with the 'roan' locus. However, these two loci could be excluded from respectively 1.7 and 2.5 Morgans of the bovine genome.
Assuntos
Antígenos de Grupos Sanguíneos/genética , Bovinos/genética , Ligação Genética , Marcadores Genéticos , Animais , Feminino , Escore Lod , Masculino , Polimorfismo Genético , Recombinação GenéticaRESUMO
A new hypervariable tandem repeat was isolated from the pig genome and characterized by DNA sequence. The use of this DNA fragment as a probe in order to follow allelic segregation and DNA fingerprinting in pigs, horses and rabbits is documented.
Assuntos
DNA/genética , Suínos/genética , Animais , Sequência de Bases , Clonagem Molecular , Sondas de DNA , Feminino , Ligação Genética , Marcadores Genéticos , Masculino , Dados de Sequência Molecular , Polimorfismo de Fragmento de Restrição , Sequências Repetitivas de Ácido NucleicoRESUMO
Eighty anti-SLA class I reagents were prepared resulting from skin graft and subcutaneous immunizations in 320 fattening pigs of the Belgian Landrace and Pietrain breeds. By means of these alloantisera seven internationally and five locally established specificities were recognized. Three of the locally assigned specificities were new: BM 36, BM 37 and BM 38. They were serologically and genetically defined. The typing battery was completed with French and Danish reagents, and correlation coefficients were calculated for the main alloantisera recognizing SLA class I alloantigens observed in the Belgian breeds. The SLA haplotype frequencies were estimated in 372 Belgian Landrace and 369 Pietrain pigs. The SLA haplotype distribution differs significantly between both breeds and the genetic distance (0.54) at the SLA system is quite high.
Assuntos
Genes MHC Classe I , Antígenos de Histocompatibilidade/genética , Suínos/imunologia , Animais , Frequência do Gene , Haplótipos , Imunização , Isoanticorpos/biossíntese , Especificidade da Espécie , Suínos/genéticaRESUMO
Captivity-induced stereotypies in bank voles (Clethrionomys glareolus) were inhibited at the age of 1, 2, 4, 6, 8 and 10 months by 0.2 mg/kg haloperidol s.c., while 35 mg/kg naloxone s.c. only inhibited them at the age of 4-6 months. Other activities were not significantly inhibited. The change in the naloxone effect was not due to desensitization as animals treated only at 6 or 8 months reacted similarly. It is suggested that the expression of such stereotypies is influenced by endogeneous brain opioid peptides during an early stage of their development, but that thereafter only motoric automatisms remain, essentially under dopaminergic control.
Assuntos
Arvicolinae/fisiologia , Haloperidol/farmacologia , Naloxona/farmacologia , Comportamento Estereotipado/efeitos dos fármacos , Envelhecimento/fisiologia , AnimaisRESUMO
1. Pig serum Po2 protein and horse Xk protein were purified by FPLC, non-denaturing 2D agarose-PAGE and 2D IPG-PAGE. 2. The separated fractions were electroblotted to poly(4-vinyl-N-methylpyridinium iodide) coated GF/C glass fiber sheets. 3. The partial amino acid sequences and amino acid compositions of different genetic variants of the proteins were determined. 4. The results proved that previously reported polymorphic serum post-albumins in each of these species were homologous to human plasma alpha 1B-glycoprotein.
Assuntos
Proteínas Sanguíneas/genética , Glicoproteínas , Imunoglobulinas , Sequência de Aminoácidos , Aminoácidos/análise , Animais , Proteínas Sanguíneas/isolamento & purificação , Cavalos , Humanos , Dados de Sequência Molecular , Polimorfismo Genético , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , SuínosRESUMO
A new variant of glucose phosphate isomerase (GPI), also known as phosphohexose isomerase (PHI), was detected in a primitive pig population.