Screening of Ferritin cDNA

At the first screening, thirty-five thousand plaques were screened. As a result, 22 positive plaques were detected. Finally, 16 positive plaques were isolated after the second screening. Two kinds of cDNA were subcloned, and each clone was named either pAMFE1-1 or pAMFE1-2. Following subcloning, the sequence of these two clones was analyzed. Both clones had full length ferritin cDNA. The DNA sequence and the amino acid sequence are shown in Figure 27.4. In Figure 27.4, N-terminal amino acid sequences of acid responsive protein are completely in accord with the sequences of the underlined parts (b). It was confirmed that this cDNA clone encoded the acid responsive protein. Alignment of the amino acid sequence of acid responsive protein with the plant ferritin sequences showed 77% identity, and the putative ferric ion biding site was also conserved. These results indicated that this cDNA was the ferritin cDNA of Acacia mangium.

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