Discussion

The nucleotide sequence of the cDNA insert was used for the deduction of the corresponding amino acid sequence. Upon nucleotide search programming and computer analysis, the clone was identified as GenBank accession number T20778. No specific protein was found to be coded for the isolated gene insert, suggesting that either a protein has not been identified for this clone, or that further analysis will lead to better identification of the protein. There were also no high similarities with other plant types so the sequence we isolated may possibly not be conserved [7]. Perhaps the clone identified does not have a characterizable protein at the moment because it is a relatively new one that is still undergoing more research. Thus, no protein was listed in the databanks.

Our recombinant plasmid was found to be approximately 5.5 kb in length, with an insert size of approximately 1.5 kb. The isolated insert had a open reading frame of approximately 378 bp. Even though the ORF was 378 base pairs in length, upon performing the annotated sequence, the first 100 base pairs did not seem to code for a specific amino acid.

Southern analysis indicated that one gene copy was present in the EcoRI, BamHI, and HindIII restriction enzyme cuts. The presence of other bands in the Southern experiment show that other genes similar to the one we isolated exist, and may be related to our clone. Northern experiments showed the gene expression to be influenced by wounding due to the presence of the 2.4 kb fragment in the 5 hour wounded sample. A lack of fragments in the tissue-specific lanes does not allow for the isolation of the protein to any specific part of the plant. Without this information, it is difficult to understand the function of the protein in the Arabidopsis plant. The hydrophilicity data shown in Figure 5 supports the idea that the majority of the amino acids in the protein are hydrophobic. The amino acids may be buried in the membrane and not on the surface. The protein may be a membrane protein, but because of the short nature of the analyzed segment, further study is required to determine if the protein is a transmembrane protein.

In conclusion, the cDNA insert has been isolated, but no specific protein has been found that the insert codes for. It seems that the protein may serve some function at the membrane of the cell, perhaps acting as a transmembrane protein. Continuing experiments and analysis will help in elucidating the function(s) and some special traits of the protein involved. Also, the availability of more data can correct for possible errors made previously in searching for homologies between known sequences and give us a more definitive conclusion as to the protein and its functional role. New information will provide more depth and insight into which genes are highly conserved between Arabidopsis thaliana and other different plant types as well as which genes are being maintained in species beyond the Plant Kingdom [6].