非同位素标记法检测人染色体端粒长度  

A NONRADIO-LABELLED METHOD ASSAYS TO MEASURE THE TELOMERE LENGTH OF HUMAN CHROMOSOME

作　　者：许丽艳[1] 李恩民[2] 沈忠英[1] 蔡唯佳[1] 沈健[1] 

机构地区：[1]汕头大学医学院病理学教研室,广东汕头515031 [2]汕头大学医学院生物化学与分子生物学教研室,广东汕头515031

出　　处：《癌变．畸变．突变》2001年第1期1-4,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金重点项目!(39830 380 )

摘　　要：目的 :建立一种非同位素标记测定端粒长度的方法。方法 :以地高辛标记寡核苷酸 (CCCTAA) 3 为探针 ,对基因组DNA进行Southern杂交 ,NBT/BCIP显色后与DNA标准分子量比较 ,图象分析系统测定端粒长度。结果 :通过严格控制探针的工作浓度 (1:12 80稀释 )、DNA上样量 (>2 0 μg)、杂交温度 (4 7± 2℃ )等主要影响因素 ,获得了比较好的杂交条带。结论 :建立了一种稳定、可靠、低背景的地高辛标记检测端粒长度的方法。Purpose: To establish a nonradio-labelled method for measuring telomere length (mean length of telomere restriction fragment, TRF) of human chromosome. Methods: TRF was examined by Southern blotting by using labelled oligonucleotide (CCCTAA)\-3 probe with digoxin. After hybridization, the blot was stained with NBT/BCIP, analysed by using a densitometric imaging system and mean length of TRF calculated, comparing with standard kilobase sized marker. Results: The hybridization bands were showed under stringently controling the prode working concentration (1:1 280 dilution), loaded DNA sample (>20 μg) and hybridization temperature (47±2℃) Conclusion: A digoxin-labelled, stable, reliable and low background method for determining the telomere length of human chromosome was established.

关 键 词：端粒 端粒长度 非同位素标记 癌变细胞 

分 类 号：R730.231[医药卫生—肿瘤] R363.14[医药卫生—临床医学]