subtractive hybridization strategy for the identification of differentially expressed genes was performed between LPSchallengedand naive Ciona intestinalis. This strategy allowed the characterization of two transcripts (Ci8short and Ci8long)generated by the use of two Alternative Polyadenylation sites. The Ci8long transcript contains a protein domain withrelevant homology to several components of the Receptor Transporting Protein (RTP) family not present in the Ci8shortmRNA. By means of Real Time PCR and Northern Blot, the Ci8short and Ci8long transcripts showed a different pattern ofgene expression with the Ci8short mRNA being strongly activated after LPS injection in the pharynx. In situ hybridizationanalysis demonstrated that the activation of the APA site also influenced the tissue localization of the Ci8short transcript.This analysis showed that the Ci8long mRNA was expressed in hemocytes meanwhile the Ci8short mRNA was highlytranscribed also in vessel endothelial cells and in the epithelium of pharynx. These findings demonstrated that regulation ofgene expression based on different polyadenylation sites is an ancestral powerful strategy influencing both the level ofexpression and tissue distribution of alternative transcripts.
|Numero di pagine||12|
|Stato di pubblicazione||Published - 2013|
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