Detection of SNP of the pain related human μ opioid receptor gene by primer-directed pyrosequency to predict the dose of analgesic

• Main Authors: Wen-Ying Chou, 周文英
• Other Authors: none
• Format: Others
• Language: en_US
• Online Access: http://ndltd.ncl.edu.tw/handle/61220374667654683404

碩士 === 高雄醫學大學 === 藥學研究所碩士在職專班 === 92 === Human mu opioid receptor (MOR) plays an essential role in mediating effects of opioids. Epidemiological studies have shown that opiate addiction has a heritable characteristic. Both clinical and laboratory studies suggest that the MOR gene may contribute to the heritable component of susceptibility to develop opiate addiction. Single nucleotide polymorphisms (SNPs) have been identified in the MOR gene by conventional methods. One of these candidate coding region SNPs, the A118G substitution, occurs at high allelic frequencies (10.5% ). The A118G substitution encodes a variant receptor with binding and signal transduction differences in response to β-endorphin in cellular assays. This common SNP causes amino acid changes in the receptor, and may have implications for decreased or increased susceptibility to opiate addiction, as well as differences in individual responses to opioids. Many papers have shown decreased or increased susceptibility to opiate addiction, we want to find the differences in individual responses to opioids here. Recent innovations in pyrosequencing technology offer a new and accurate method for SNP detection. We suggest here to use pyrosequencing technology for detection of the SNP of the MOR gene Taiwanese samples. First, PCR-amplified genomic DNA samples are used to produce target fragments. The target fragments will be used as templates for analysis by Pyrosequencing with a sequencing primer 2 bp upstream of the mutation spot. In this project, all 100 samples will be sequenced by Pyrosequencing for genotypes of normal homozygous, heterozygous, and mutant homozygous. Genotypes will be determined and allelic frequency calculated for correlation between MOR SNP and opiate for analgesic.