JIANG Tingbo;LI Fengjuan;YANG Chuanping

;Heilongjiang Key Laboratory of Forest Tree Genetic Improvement, Northeast Forestry University, Harbin Heilongjiang 150040, PRC

ferritin;gene expression;tobacco;real-time rt-pcr

To understand the use of real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) for detecting the relative abundance of mRNA, the expression of a tobacco ferritin gene (A/tFer1) was detected by Northern blot and real-time RT-PCR.The results indicated that both of the two methods were able to detect mRNA expression of A/tFer1 clearly and similarly, namely NtFert expression was responsive to iron-overload, and the abundance of A/tFer1 mRNA was greatly increased after iron loadedfor 6 h. To compare the effect and sensitivity of two methods, results revealed that Northern blot need 30 mu g of total RNA and at least 3 days for the total protocol performance, whereas real-time RT-PCR only need 2 jxg of total RNA and 1.5 h. The real-time RT-PCR is rather sensitive and effective than Northern blot. Real-time RT-PCR analysis can be used to rapidly detect the relative abundance of mRNA expression instead of Northern blot analysis.