RESUMO
One hundred and seventy-four rat loci which contain short tandem repeat sequences were extracted from the GenBank or EMBL data bases and used to define primers for amplification by the polymerase chain reaction (PCR) of the microsatellite regions, creating PCR-formatted sequence-tagged microsatellite sites (STMSs). One hundred and thirty-four STMSs for 118 loci, including 6 randomly cloned STMSs, were characterized: (i) PCR-analyzed loci were assigned to specific chromosomes using a panel of rat x mouse somatic cell hybrid clones. (ii) Length variation of the STMSs among 8 inbred rat strains could be visualized at 85 of 107 loci examined (79.4%). (iii) A genetic map, integrating biochemical, coat color, mutant and restriction fragment length polymorphism loci, was constructed based on the segregation of 125 polymorphic markers in seven rat backcrosses and in two F2 crosses. Twenty four linkage groups were identified, all of which were assigned to a defined chromosome. As a reflection of the bias for coding sequences in the public data bases, the STMSs described herein are often associated with genes. Hence, the genetic map we report coincides with a gene map. The corresponding map locations of the homologous mouse and human genes are also listed for comparative mapping purposes.
Assuntos
Mapeamento Cromossômico/métodos , DNA Satélite/genética , Reação em Cadeia da Polimerase , Animais , Sequência de Bases , DNA de Cadeia Simples , Bases de Dados Factuais , Ligação Genética , Células Híbridas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Polimorfismo Genético , Ratos , Sequências Repetitivas de Ácido NucleicoRESUMO
A genomic renin exon 9 fragment was subcloned into vector pSPT18 and used for in vitro transcription to obtain 32P-labeled rat renin cRNA. Using this cRNA, we established quantitative solution hybridization and specific Northern blotting assays for renin mRNA. We were able to detect renin mRNA in the kidney, heart ventricle and atrium, brain, testis, and adrenal gland of male rats in the concentrations of 430 +/- 8.1, 110 +/- 1.9, 43 +/- 0.9, 64 +/- 1.1, 47 +/- 0.9 and 11 +/- 0.2 pg mRNA/mg of total RNA, respectively.
Assuntos
Vetores Genéticos , Hibridização Genética , RNA de Cadeia Dupla/genética , RNA Mensageiro/análise , Renina/genética , Animais , Clonagem Molecular , Éxons , Rim/análise , Masculino , Miocárdio/análise , RNA Mensageiro/genética , Ratos , Ratos Endogâmicos WKY , Renina/análise , Testículo/análise , Transcrição GênicaRESUMO
The complexity of recognizing the potential contribution of a number of possible predictors of complex disorders is increasingly challenging with the application of large-scale single nucleotide polymorphism (SNP) typing. In the search for putative genetic factors predisposing to coronary artery restenosis following balloon angioplasty, we determined genotypes for 94 SNPs representing 62 candidate genes, in a prospectively assembled cohort of 342 cases and 437 controls. Using a customized coupled-logistic regression procedure accounting for both additive and interactive effects, we identified seven SNPs in seven genes that, together, showed a statistically significant association with restenosis incidence (P <0.0001), accounting for 11.6% of overall variance observed. Among them are candidate genes for cardiovascular pathophysiology (apolipoprotein-species and NOS), inflammatory response (TNF receptor and CD14), and cell-cycle control (p53 and p53-associated protein). Our results emphasize the need to account for complex multi-gene influences and interactions when assessing the molecular pathology of multifactorial medical entities.