The NOD2 gene codes for an important protein in the innate immune response to bacteria of the human body. Up until recent years, the RNA sequence has been sequenced only for lymphatic cells, but only alternative isoforms are expressed in epithelial cells of the colon and small intestine. The alternative isoform 1a-2 is found more often in the colon and small intestinal cells of people suffering from the chronic disease Crohn’s. The alternative isoforms have not yet been fully sequenced, only the first 3 exons have been sequenced. The goal of this paper is to analyze and sequence the 3’ end of the mRNA in epithelial cells, with a basis in the DQ868973 sequence that codes for the 1a/2 isoform. This was done by replacing the first 60 base pairs of the conventional NOD2 mRNA RefSeq, with the base pairs from the DQ868973 mRNA sequence. We designed the primers on this basis for the expected alternative isoform, and used an Oligo(DT) primer for cDNA synthesis. The expected cDNA fragment with our self-designed primer set 1 was 3913 base pairs long binding at base pair 23-41 for the forward primer, and 3935-3916 for the reverse primer. We did not manage to sequence the gene, but we ran a gel which showed promise that the DQ868973 isoform give way to an mRNA sequence at least, if not more than, 3913 base pairs long.
|Uddannelser||Basis - Naturvidenskabelig Bacheloruddannelse, (Bachelor uddannelse) Basis|
|Udgivelsesdato||22 jun. 2015|