实验动物恒河猴朊病毒基因分析

Analysis of Prion Genes in Rhesuses

目的 通过对拟应用于航天飞船模拟试验的实验动物恒河猴进行朊病毒基因检测和分析 ,了解其朊病毒疾病易感性 ,进而为该试验所用实验动物质量提供朊病毒疾病的遗传数据。方法 根据已报道的猴朊病毒基因序列设计引物对 ,采用PCR方法扩增恒河猴朊病毒基因 ,将其克隆到T—VECTOR ,序列测定及分析。结果 序列测定及分析表明所克隆的恒河猴PRNP基因片段为 76 2bp ,该基因内无内含子 ,包含了PRNP完整编码区序列 ,编码 2 5 3个氨基酸的前体蛋白 ,推测其相对分子质量约 2 7 6× 10 3。与已报道的猴的相应序列作比较 ,核苷酸序列同源性分别为 95 %以上 ,其编码的氨基酸同源性均为 95 %以上。其中发现了已报道的多态性位点N97S ,H10 0N和未曾见报道的Q5 2E点突变位点 ,以及沉默突变位点A78G ,T84C ,T4 95C ,A5 6 4G ,C6 5 4 ,未发现与朊病毒敏感性连锁的氨基酸多态位点P10 2L、M12 9V ,N171S和E2 19K。结论 航天医学实验所用 17只恒河猴不存在朊病毒敏感性基因。

Objective To analyze whether there are sensitive PRNP genes in the r hesuses used in spaceship analog experiments, and to provide a reference for ass essment of pathological brain alterations of these rhesuses tprovoked by exposur e to hypergravity.Method\ Total DNA was isolated from peripheral bl ood ymphocyte s of 17 healthy male adult rhesuses, which would be used in corresponding parabo lic flight experiments. The PRNP gene was amplified by PCR and analyzed.R esults\ The cloned PRNP gene is 762 nucleotides long and contains the en tire PRNP coding sequence, which has no intron and has more than 95% homology w ith the published gene sequence, deducing 253 amino acids and molecular weight ( MW) of 27.6 kd.Conclusion\ There are no sensitive PRNP genes in th e rhesuses, which provide a reference for judgement of pathological brain alterations of these rhesuses as e xposed to hypergravity.