摘要
目的 建立荧光定量聚合酶链反应 ( fluorogenic quantitative PCR)检测方法 ,以便快速、准确地定量检测细胞株转录因子表达强度。方法 根据基因序列 ,设计合成引物 ,采用一步法提取细胞株总 RNA,逆转录获取 c DNA,通过荧光定量的方法对各细胞株的转录因子 T-box expression in T cells( T-bet)、GATA3的表达水平进行检测。结果 细胞株NK92、QBC93 9、K5 62、HO-891 0、A5 49、He La转录因子 T-bet的表达强度分别为 0 .90、1 .2 5、0 .96、0 .92、1 .1 1和 1 .1 7,GA-TA3的表达强度分别为 1 .2 3、1 .49、1 .3 0、0 .97、1 .0 5和 1 .3 4。结论 建立了肿瘤细胞株转录因子基因表达的荧光定量PCR检测方法 ,较常规 PCR方法更为简便、快速、准确 ,有很好的应用前景。
Objective To establish fluorogenic quantitative PCR method for detecting the gene expression of transcription factors. Method According to the specific sequence from gene bank,the primers were designed, total RNA were obtained by Trizol, then cDNA were synthesized,the gene expression of transcription factors T-bet,GATA3 were detected by fluorogenic quantitative PCR. Results The expression of transcription factors T-bet in NK92,QBC939,K562,HO-8910,A549,HeLa were 0.90,1.25,0.96,0.92,1.11 and 1.17 respectively,while GATA3 were 1.23,1.49,1.30,0.97,1.05 and 1.34.Conclusion Fluorogenic quantitative PCR for the detection of the gene expression of transcription factors has been established successfully.
出处
《临床输血与检验》
CAS
2004年第2期86-88,共3页
Journal of Clinical Transfusion and Laboratory Medicine
基金
安徽省自然科学基金资助 ( No.0 3 0 43 70 3 )
