Quantitative Detection of P53 mRNA by Molecular Beacons in vitro

Abstract

A p53 molecular beacon (p53 MB) designed according to the p53 gene sequence was synthesized to quantitatively detect p53 mRNA in vitro. The specificity of the p53 MB was tested by hybridizing the MB with total RNA extracted from CNE2, CNE2-vector (CNE2 cells transfected with empty plasmid) and CNE2-p53 RNA interference (CNE2 cells transfected with p53 RNA interference plasmid) cell lines. Lower fluorescence intensity in CNE2-p53 RNA interference cell samples confirmed the specificity of the p53 MB to the p53 mRNA. The change of the p53 mRNA expression in various kinds of cell lines was measured by traditional RT- Polymerase Chain Reaction (PCR). Based on this, the p53 mRNA expression in the lung carcinoma cells of A549 treated with 5-fluorouracil at various time was detected by p53 MB in vitro. These results indicated the rapid response of p53 MB to the effect of the drug on the expression of p53 mRNA in cells.

Key words: molecular beacon, p53 RNA interference, 5-fluorouracil, quantitative

Cite this article

HE Li-Fang^1,2, TANG Hong-Xing^1,2, WANG Ke-Min*^,1,2,c,TAN Wei-Hong^1,2,c, LIU Bin^2,c, MENG Xiang-Xian^2,c, LI Jun^1,2,c, WANG Wei^{1,. Quantitative Detection of P53 mRNA by Molecular Beacons in vitro[J]. Acta Chimica Sinica, 2006, 64(11): 1116-1120.}

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分子信标用于p53 mRNA的体外定量检测