RESUMO
A population-based study was undertaken to evaluate linkage between single-nucleotide polymorphisms known as risk factors and type 2 diabetes in an Indian population. The study population was comprised of 40 normal glucose-tolerant individuals (21 males and 19 females) and 40 type 2 diabetes patients (21 males and 19 females). The genes and their corresponding single-nucleotide polymorphisms that we screened were VDR (rs 731236 and rs 1544410), IL-6 (rs 1800795), TCF7L2 (rs 7903146) and TNF-α (rs 1800629). The risk alleles were more frequent in the subjects with type 2 diabetes, except for the TNF-α gene, which was very infrequent in the population; the normal allele occurred at high and similar frequencies in both normal and diabetic individuals.
Assuntos
Diabetes Mellitus Tipo 2/genética , Genômica , Polimorfismo de Nucleotídeo Único , Adulto , Feminino , Humanos , Índia , Masculino , Pessoa de Meia-IdadeRESUMO
The technology of mRNA-based differential display reverse transcriptase-polymerase chain reaction (DDRT-PCR) was used to detect a 246-bp differentially expressed fragment from the nematophagous fungus Arthrobotrys oviformis when young mycelia were induced with the round worm Haemonchus contortus. The fragment was converted into an expressed sequence tag (EST) through characterization at the molecular level. Homology search indicated that the differentially expressed fragment originated from the cuticle-degrading serine protease gene, which has been previously reported to play a role in nematophagous activity in A. oligospora, Dactylaria parvispora, A. musiformis, and other potential anti-fungal biological control agents. Several single nucleotide polymorphisms found to represent both synonymous as well as non-synonymous mutations within this short sequence stretch of 246 bp suggested genetic variability within the gene in this group of nematode-trapping fungi. The cloned EST fragment has potential for use as a hybridization probe for searching full-length gene from an appropriate cDNA library of this and related fungi. This is the first report of the identification of an EST representing the cuticle-degrading serine protease gene from A. oviformis using the technique of DDRT-PCR.
Assuntos
Ascomicetos/enzimologia , Ascomicetos/genética , Etiquetas de Sequências Expressas , Proteínas Fúngicas/genética , Perfilação da Expressão Gênica/métodos , Serina Endopeptidases/genética , Sequência de Aminoácidos , Ascomicetos/metabolismo , Sequência de Bases , Proteínas Fúngicas/metabolismo , Genes Fúngicos/genética , Dados de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Serina Endopeptidases/metabolismoRESUMO
An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle. This has opened the prospect for large-scale screening of cattle using the polymerase chain reaction (PCR) technique for the rapid identification of heterozygous animals. To facilitate such a screening process, the mutant and normal alleles of factor XI gene, represented by 244- and 320-bp PCR amplified fragments, were individually cloned in Escherichia coli using a multicopy plasmid cloning vehicle to generate pFXI-N and pFXI-M, respectively. The authenticity of the inserts was confirmed by nucleotide sequencing. A nested PCR method was developed, by which PCR amplicons generated from primers with annealing sites on the recombinant plasmids and by flanking the insert were used as templates for amplification of the diagnostic products using factor XI gene-specific primers. An equimolar mixture of both PCR amplicons, originating from pFXI-N and pFXI-M, constituted the carrier control while the individual amplicons were the affected and normal controls. The controls were used as references for in-gel comparison to screen a population of 307 cattle and 259 water buffaloes; the frequency of the mutant allele was found to be 0. No DNA size standards were required in this study. The simulated control DNA samples representing normal, carrier and affected cattle have the potential to help in large-scale screening of a cattle population for individuals that are carriers or affected by factor XI deficiency.
Assuntos
Doenças dos Bovinos/genética , Deficiência do Fator XI/veterinária , Triagem de Portadores Genéticos/métodos , Análise de Sequência de DNA/veterinária , Alelos , Animais , Sequência de Bases , Búfalos , Bovinos , Deficiência do Fator XI/genética , Genótipo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/veterináriaRESUMO
An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle. This has opened the prospect for large-scale screening of cattle using the polymerase chain reaction (PCR) technique for the rapid identification of heterozygous animals. To facilitate such a screening process, the mutant and normal alleles of factor XI gene, represented by 244- and 320-bp PCR amplified fragments, were individually cloned in Escherichia coli using a multicopy plasmid cloning vehicle to generate pFXI-N and pFXI-M, respectively. The authenticity of the inserts was confirmed by nucleotide sequencing. A nested PCR method was developed, by which PCR amplicons generated from primers with annealing sites on the recombinant plasmids and by flanking the insert were used as templates for amplification of the diagnostic products using factor XI gene-specific primers. An equimolar mixture of both PCR amplicons, originating from pFXI-N and pFXI-M, constituted the carrier control while the individual amplicons were the affected and normal controls. The controls were used as references for in-gel comparison to screen a population of 307 cattle and 259 water buffaloes; the frequency of the mutant allele was found to be 0. No DNA size standards were required in this study. The simulated control DNA samples representing normal, carrier and affected cattle have the potential to help in large-scale screening of a cattle population for individuals that are carriers or affected by factor XI deficiency.