影响启动子活性测定的关键影响因素

目的探讨载体、质粒转染剂量、启动子长度、细胞系以及检测仪器等因素对启动子分析实验的影响。方法采用化学发光法测定荧光素酶,EGFP荧光强度采用流式细胞仪。以上检测每组样品均设三复孔,并重复两次,所得结果进行t检验。结果选择不同的报告基因、载体、质粒转染剂量、启动子长度和细胞系得到不同的结果。结论载体、质粒转染剂量、启动子长度、细胞系的选择会对启动子分析的结果造成很大的影响,因此,在进行启动子分析前,应综合考虑上述因素,以达到更好的实验效果。

Experience on Analyzing the Promoter Activity with Luciferase and Green Florence Protein

Objective To investigate the effect of vector,dose,inner reference,length of the promoter,cell line and the instrument to promoter analysis.Method Determine the luciferase with chemiluminescence and the fluorescence intensity of EGFP with flow cytometer.Each specimen be tested in three action-well and the tests were repeated twice.The result be checked with t-test.Result It would get different result when choose different reporter gene,vector,dose,enner reference,length of the promoter,cell line,and the instrument.Conclusion The vector,dose,inner reference,length of the promoter,cell line and the instrument will make great effect to the result of promoter analysis.Therefore,to get preferably result it is important to think the factor synthetic before the promoter analysis be done.