人内源性逆转录病毒（HERV）序列在人星形胶质细胞系及运动神经元病患者血清中的检测与分析

发布时间：2018-01-02 11:42

本文关键词：人内源性逆转录病毒（HERV）序列在人星形胶质细胞系及运动神经元病患者血清中的检测与分析　出处：《吉林大学》2010年硕士论文　论文类型：学位论文

【摘要】： 内源性逆转录病毒(human endogenous retrovirus,HERV)是大约几百万年前整合到人类祖先生殖细胞中的逆转录病毒,有30-50个家族,占人类基因组DNA含量的8%。在人类进化过程中,由于突变、缺失、插入等作用,大多数HERV已经不具备编码能力,但是仍有一部分HERV依然有完整的开放阅读框,甚至能形成逆转录病毒样颗粒。近年来研究发现,HERV可能与自身免疫性疾病,如类风湿性关节炎;一些肿瘤,如人黑色素瘤、睾丸生殖细胞瘤、绒毛膜癌、畸胎瘤等;神经退行性疾病,如多发性硬化症、肌萎缩性脊髓侧索硬化等很多种疾病相关。本实验首先以人星形胶质细胞系为研究对象,检测HERV的拷贝数,又利用运动神经元病(motor neuron disease,MND)患者血清及非MND患者血清,探讨HERV与MND的相关性。分别提取细胞及患者血清中DNA,进行实时定量PCR检测HERV-K pol在人星形胶质细胞及患者血清中的拷贝数;利用常规PCR产物进行TA-cloning,最终进行序列分析、比对,以检测HERV-K pol序列在人星形胶质细胞系及MND患者血清中存在的基序突变,从而判断HERV-K pol基因突变与MND发病机制的相关性。为进一步研究HERV与脑肿瘤及MND的关系提供线索。
[Abstract]:Endogenous retrovirus (human endogenous, retrovirus, HERV) is about millions of years ago to integrate human ancestors in germ cells of the retrovirus, 30-50 family, accounting for the human genome content of DNA 8%. in human evolution, due to mutation, deletion, insertion and so on, most of the HERV does not have the encoding ability, but there are still a part of the HERV still has a complete open reading frame, even the formation of retrovirus like particles. In recent years, the study found that HERV may be related to autoimmune diseases, such as rheumatoid arthritis; some tumors, such as melanoma, germ cell tumor, testicular choriocarcinoma, teratoma; neurodegenerative diseases, such as multiple sclerosis, amyotrophic lateral sclerosis and other diseases.
The first experiment in human astrocyte cell line as the research object, the copy number detection of HERV, and the use of motor neuron disease (motor neuron, disease, MND) in serum of patients with serum and non MND, to investigate the correlation between HERV and MND. Were extracted from the cells and serum in patients with DNA, the copy number for detection of HERV-K by real-time pol PCR in astrocytes and in the serum of the patients; the use of TA-cloning product of conventional PCR, the final sequence analysis, alignment, motif for detection of HERV-K pol sequences present in the serum of patients with human astrocyte cell line and MND mutation in HERV-K, in order to determine the relationship between pol gene mutation and the pathogenesis of MND provide clues. Further study the relationship between HERV and brain tumors and MND.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R346

【共引文献】