红白血病K562细胞基因表达谱芯片制作研究
K562细胞,,K562细胞；基因表达谱；DNA,微集阵列,1材料和方法,2结果,3讨论,参考文献
摘 要 以人红白血病K562细胞为材料，应用限制性显示PCR(RD-PCR)技术分离到K562细胞在诱导分化药物羟基脲诱导分化前后不同时期的cDNA片段3256条，并用DNA芯片仪制作了K562细胞基因表达谱芯片。对芯片制作中不同点样液及处理条件对玻片上DNA固定效率的影响，样品DNA最优浓度等进行了初步研究，结果认为用DMSO作点样液、样品DNA浓度为0.3 μg/μL、点样后经紫外交联(150 mJ)、80 ℃干烤2 h, DNA探针在玻片上的固定效率可达95%。用此法制作的K562基因表达谱芯片得到较好的杂交效果。关键词 K562细胞；基因表达谱；DNA 微集阵列
Fabrication of cDNA Microarray for the Gene Expression in Leukemia K562 Cell Line
ZHU Ji1,2， MA Wenli1, SONG Yanbin1, WU Qinghua2, YAO Ruhua2, ZHENG Wenling3
(1.Institute of Mol. Bio., First Military Medical University, Guangzhou,Guangdong 510515; 2. Department of Bioeng., South China University of Technology; 3. General Hospital of Guangzhou Military Command)
Abstract 3256 cDNA fragments were isolated from human leukemia K562 cells, induced with hydroxyurea by restriction display PCR(RD-PCR). DNA microarray of K562 cell gene expression profile was prepared. Influences of different spot solutions and treatments on DNA attachment and optimal DNA concentration were studied. The results showed that DNA attachment rate was 95% when DMSO was used as spot solution, DNA concentration was 0.3 μg/μL, microarray was crosslinked by UV ray(150 mJ) and dried at 80 ℃ for 2 h. A good crosslinking effect was achieved using the DNA microarray of K562 cell gene expression profile prepared in this study. ......
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