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本研究旨在探讨穿孔素(perforin)基因PRF1突变是否为获得性重型再生障碍性贫血(SAA)发病的遗传易感性基础。用聚合酶链反应(PCR)方法扩增31例SAA患者及15名正常对照者外周血单个核细胞基因组DNAPRF1外显子2(exon2)及外显子3(exon3);用双脱氧终止法测序,与GenBank报道序列核对寻找突变位点;发现突变序列克隆入M13噬菌体载体中,对所得2条染色体相应序列分别测序,以确定不同突变在染色体上的分布。结果表明:在SAA患者,发现了2处基因突变即822位C>T纯合子突变(无义突变)及907位G>A杂合子突变(Met303Val);rs885822单核苷酸多态性(SNP)位点分布病例组与对照组比较有统计学差异(p<0.05)。结论:穿孔素基因突变可能是部分SAA患者的遗传易感因素。
The purpose of this study was to investigate whether the PRF1 mutation in the perforin gene is the underlying basis of genetic susceptibility to acquired severe aplastic anemia (SAA). Polymerase chain reaction (PCR) was performed to amplify exon2 and exon3 of PRF1 gene in peripheral blood mononuclear cells from 31 patients with SAA and 15 healthy controls. Sequencing by dideoxy terminator , And the sequence of GenBank was checked to find the mutation site. The mutation sequence was cloned into the M13 phage vector and the corresponding sequences of the two chromosomes were sequenced separately to determine the distribution of the different mutations on the chromosome. The results showed that in the SAA patients, two genetic mutations were found, namely 822 C> T homozygous mutation (nonsense mutation) and 907 G> A heterozygous mutation (Met303Val); rs885822 single nucleotide polymorphism ) Site distribution of cases and control groups were statistically significant (p <0.05). Conclusion: Perforin gene mutation may be a predisposing factor in some SAA patients.