【作者】 杨建民； 赵德明； 阚广捍； 李宁； 郝永新； 李莹辉； 吴斌；

【Author】 YANG Jian-min ,ZHAO De-ming ,KAN Guang-han ,LI Ning , HAO Yong-xin , LI Ying-hui . WU Bin( College of Veterinary Medicine, National Laboratory for Animal Transmissible Spongiform Encephalopathy (TSE) ,China Agriculture University, Beijing 100094,China; Space Medicine (Medical Engineering, Beijing 100094,China;National Laboratory for Agro biotechnology, China Agriculture University, Beijing 100094,China)

【机构】 中国农业大学动物医学院； 航天医学工程研究所； 中国农业大学农业生物技术国家重点实验室；

【摘要】 目的通过对航天医学研究飞船应急返回高过载模拟试验中所用实验动物恒河猴进行朊病毒基因诊断和分析,了解实验动物的朊病毒疾病易感性,进而为该试验所用实验动物质量提供背景资料和朊病毒的检测数据。方法根据已报道的猴朊病毒基因序列设计引物对,采用PCR方法扩增了应用于航天模拟试验的恒河猴朊病毒基因,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762bp,该基因内无内含子,包含了PRNP完整编码区序列,编码253个氨基酸的前体蛋白,推测其分子量约27.6kd。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,H100N和未曾见报道的Q52E点突变位点,以及沉默突变位点A78G,T84C,T495C,A564G,C654,未发现与朊病毒敏感性连锁的氨基酸多态位点P102L、M129V,N171S和E219K 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。这为航天医学研究飞船应急返回高过载模拟试验中所用动物质量提供了朊病毒检测的依据,进而为实验动物各项生理病理指标的测定,尤其在研究高过载模拟试验条件下对动物脑部变化的影响提供了很有价值的参考数据。更多还原

【Abstract】 Objective To investigate whether there are sensitive PRNP genes in the rhesuses being used in the spaceship analog experiment, so as to provide reference to judge the brain pathology of these rhesus to high G. Methods The total DNA was isolated from lymphocyte of peripheral blood of Seventeen healthy male adult rhesuses, which would be used in the corresponding parabolic G curve experiments. The PRNP gene was amplified by PCR and further analyzed. Results The cloned PRNP gene was 762-nucleotide long and contained the entire PRNP coding sequence, which has no intron and has more than 95% homology with the published gene sequence, and was translwted into 253 amino acids which had a calculated molecular weight ( MW) of 27.6 kd. Conclusion There are no sensitive PRNP genes in the rhesuses, and which provide reference to judge the brain pathology of these rhesuses to high G.更多还原