46，XX，t（5；15）（q21.1；q21.2）相互易位染色体断裂重接点定位方法探索

发布时间：2018-08-05 10:42

【摘要】：背景：染色体相互易位存在染色体的断裂重接,可通过改变基因内部结构或基因与其作用元件相对位置,从而导致基因表达异常而表现出疾病表型。常规细胞遗传学检测只能对断裂区域作出粗略估计,无法精确定位。新发展起来的Array painting及多种基于第二代测序技术的定位方法对实验人员的数据分析能力要求严格,且费用昂贵。目的：运用细胞遗传学、分子细胞遗传学、分子遗传学等相关技术,精确定位一例有异常表型的46,XX,t(5；15)(q21.1；q21.2),15pss相互易位患者的染色体断裂点。建立染色体断裂点定位的常规方案,探讨染色体重排与表型的关系。方法：本实验从三条技术路径尝试对染色体断裂点进行精确定位：1、短片段探针荧光原位杂交定位法；2、长片段PCR定位法；3、激光显微切割与PCR结合定位法。结果：方法1和方法2未能得到理想结果,运用方法3将断裂点定位在小于2kb范围内。结论：激光显微切割与PCR结合定位法可运用于对染色体断裂点的精确定位。
[Abstract]:Background: chromosomal translocation may result in abnormal gene expression by altering the internal structure of the gene or the relative position of the gene to its acting elements. Conventional cytogenetic testing can only make a rough estimate of the fracture area and cannot accurately locate it. The newly developed Array painting and many localization methods based on the second generation sequencing technology require strict and expensive data analysis ability of the experimenters. Objective: to accurately locate the chromosomal breakpoints in a patient with abnormal phenotype by cytogenetics, molecular genetics and other related techniques. To study the relationship between chromosome rearrangement and phenotype, a routine scheme of chromosome breakpoint location was established to study the relationship between chromosome rearrangement and phenotype. Methods: in this experiment, we try to locate the chromosome breakpoints precisely at 1: 1 by three technical paths, by fluorescence in situ hybridization (Fish) with short fragment probes, 3 by long fragment PCR, and by the combination of laser microdissection and PCR. Results: methods 1 and 2 failed to obtain ideal results. Method 3 was used to locate the fracture point in the range of less than 2kb. Conclusion: laser microdissection combined with PCR can be used to locate chromosome breakpoints accurately.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R346

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