急性白血病线粒体dna+d-loop区突变的研究

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1、目录中文摘要···············································································································英文摘要……···············································································································符号说明……︸一。前︸··················································

2、·······················································材料与方法……········································，····································································果……论······················································，，·····················································……论········

3、·····································································································……·············································································································录……参考文献············································································

4、·································……致谢︸···················································，·····················攻读学位期间发表的学术论文……山东大学硕十学位论文一急性白血病线粒体区突变的研究硕士研究生姜玉杰导师丛雅琴教授专业内科血液病中文摘要研究目的线粒体是高等动物细胞内除细胞核外唯一含有的细胞器，且能不依赖核基因组进行复制、转录和翻译，是进行氧化磷酸化反应。而理并为细胞提供能量的主要场所环区一区是线粒体的主要非编码区，占全部的左右，位置在，富含一碱基，遗传一和之间上是高变

5、，，区的重链和轻链复制起始点均位于此区内主要是调控的转录和复制，研究表明一环的突变速率约为整个序列的倍。环区具有高度，，多态性这种高度多态性反映了线粒体的高度突变性已经有大量的研究工作证实，该区是突变的热点区域。近年来，对于人类的突变研究成为，医学界尤其是肿瘤的发生发展的研究热点但对于急性白血病患者一区的突变情况尚未见有报道。本研究的主要目的是，利用技术及载体克隆技术提取并扩增急性白血病患者骨髓单个核细胞中一区，一，片段全长用直接测序法测定急性白血病患者区序列探。一，索其有无突变及突变类型并总结急性白血病患者区的突变规律探讨其发生的可能机制及后果，为急性白血病的发病机制、诊断及治疗提

6、供分子生物学方面的理论参考。研究方法选择例急性白血病患者的经抗凝的骨髓液，首先以淋，一巴细胞分离液用差速离心法分离骨髓单个核细胞用酚氯仿抽提法提取基因组，设计用于扩增线粒体一区全长的引物，上游引物为山东大学硕士学位论文，一一’一下游引物为’一一’一。用方法将其线粒体一区扩增，反应条件为起始变性温度为“，接着进行个循环为变性温度“，退火温度“，“延伸，结束延伸温度“。再用低溶点胶法回收产物，最后用载体克隆纯化产物并将含目的片段的菌液送测序，最后运用美国数据库的基因序列分析，一软件将所测得的急性白血病患者区序列进行核普酸序列同源性分析，确定所检测的序列与人类线粒体同源，然后用软件将所检测

7、序列与基因库数据对照后找出不同的位点，再与一，数据库里记录的人类线粒体区多态性变化相比较并总结起其突变类型和规律。、一，结果一成功扩增出区全长序列在处出现明显。、，一条带二例急性白血病患者中发现有例患者骨髓细胞，。，区发生了突变共发现个单核昔酸变化其中发生在高变区的有，，一个占个与中记载的区多态现象，，，。一一致其余个为碱基置换个为插入个为缺失其中均为不或之间的碱基置换，在这些置换中，主要为同质性突变，占突变总数的，其中大多数为一转换，占异质性突变则占，一