Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.13.12.5 (aequorin)
 1,451 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Toll-like receptors (TLRs) play a fundamental role in pathogen recognition and activation of innate immunity. Genetic variations in TLR have been associated with reduced host immune response to TLR ligands. We developed a rapid, simple and cost-effective method for identification of two common single-nucleotide polymorphisms (SNPs) within TLR4 gene in a high-throughput format. The method consists of a single polymerase chain reaction of the region spanning the A896G and C1196T polymorphic sites, followed by two primer extension reactions at each site using primers that carry a (dA)(24) segment at the 5' end. A biotinylated nucleotide is incorporated in the extended primer. The products are captured in microtiter wells coated with streptavidin and detected using a (dT)(30)-conjugated photoprotein aequorin. A total of 209 individuals were genotyped for each SNP. The A896G and C1196T polymorphisms were found to be in linkage disequilibrium; 186 individuals (89%) were wild-type homozygous (A/A or C/C), 22 (10.5%) were heterozygotes (A/G or C/T), and 1 (0.5%) was homozygous for the mutation (G/G or T/T). The accuracy of this method was confirmed by sequencing. The newly developed method may be useful for association studies of these two SNPs with several diseases.
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PMID:High-throughput microtiter well-based bioluminometric genotyping of two single-nucleotide polymorphisms in the toll-like receptor-4 gene. 1834 60

We developed a rapid, simple, cost-effective and high sample-throughput method for the simultaneous detection of four alleles in single-nucleotide polymorphisms (SNPs). The method was applied to the simultaneous genotyping of two common SNPs within the TLR4 gene, the A896G and C1196T polymorphisms. The method consists of a single PCR of the region spanning the A896G and C1196T polymorphic sites, followed by a quadruple primer extension (PEXT) reaction in a single tube. A biotinylated nucleotide is incorporated in the extended primer. All four products are captured in streptavidin (SA)-coated microtiter wells and detected with a combination of four reporters, the photoprotein aequorin (AEQ) and the enzymes alkaline phosphatase (ALP), beta-galactosidase (GAL) and horseradish peroxidase (HRP). For each SNP, 46 individuals were genotyped. The accuracy of this method was confirmed by sequencing. The proposed quadruple-allele chemiluminometric assay provides an accurate, simple, rapid, reproducible and cost-effective method for high sample-throughput genotyping of single-nucleotide polymorphisms.
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PMID:Quadruple-allele chemiluminometric assay for simultaneous genotyping of two single-nucleotide polymorphisms. 1930 22