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DNA from fern species
Actually, I have purified a protein from plant material
of a true fern. From the pure protein a lot of its sequence information has been obtained after tryptic digestion and N-terminal sequence analysis of the resulting peptides (about 70-80 % of the proteins amino acid sequence has been determined). By use of these amino acid sequences degenerative primers have been deduced and pairs of primers from different peptides have been used for PCR together with genomic DNA (after hundreds of runs no meaningful PCR products have been obtained). Until mow, the overexpression of only one recombinant fern protein is described in literature and RNA was used (instead of DNA) for construction of a cDNA library following PCR with degenerative primers. Does someone know is there a special intron or exon problem with genomic DNA from fern species ? Is the codon usage of this "fossil" plant family much different to angiosperms or other organisms and the reason for the problems ? Any hints, help or comments to this problem is welcome. Best regards, Andreas Giessauf, PhD Institut für Chemie Universität Graz |
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