论文部分内容阅读
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 a fluorogenic quantitative polymerase chain reaction (PCR) method to detect the expression of transcription factors in cell lines rapidly and accurately. Methods According to the sequence of the gene, the synthetic primers were designed and synthesized. The total RNA was extracted by one-step method and the c DNA was obtained by reverse transcription. The expression of T-box in T cells (T-bet), GATA3 The level of expression was detected. Results The expression intensity of T-bet of NK cell line NK92, QBC93 9, K562, HO-891 0, A549 and He La were 0 .90, 1.25, 0.96, 0.92, The expression intensities of GA-TA3 were 1 2 3,1 .49, 1 .3 0,0 .97, 1 .0 5 and 1 .3 4, respectively. Conclusion The method of real-time fluorescence quantitative PCR for gene expression of transcription factors in tumor cell lines is established. Compared with conventional PCR methods, it is simple, rapid, accurate and has a good prospect of application.