Isolating and characterizing a cDNA encoding the iron-storage protein in rice
2003
Kang. K.K. | Cho, Y.G.
A cDNA fragment encoding the iron-storage protein (ferritin) was isolated by reverse transcription (RT)-polymerase chain reaction (PCR) and plaque hybridization. The cDNA clone contained a 5' untranslated region (UTR) of 51 bp. An open reading frame of 750 bp downstream of the 5' UTR was identified, followed by a 3' UTR of 272 bp. The deduced amino acid sequence of rice protein showed 85%, 48%, and 36% identity with soybean (SolC), human (HuL-H), and horse spleen (HoS-L) ferritin, respectively. Forty-five rice varieties were evaluated for iron content in rice grain and those with the highest and lowest content were selected. The Korean rice variety Damagum showed the lowest (77.8 ppm) iron content, and Donganbyeo had the highest (234.0 ppm). The expression level of the ferritin gene was analyzed by Northern hybridization. Gene expression in Damagum was highest during the first 12-72 h after treatment of 500 mM Fe-EDTA on 21-d-old seedlings, after which it decreased. There was no expression detected after 148 h. However, the ferritin gene was highly expressed from 72 to 120 h in Donganbyeo, and the expression continued up to 172 h. The gene expression level of Donganbyeo was three times higher than that of Damagum. The possibility of transforming the rice ferritin gene is discussed.
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