Molecular analysis of serine/arginine rich SC35 splicing factor from olive

Abstract

Olive (Olea europaeaL.) is an evergreen fruit tree adapted toMediterranean climate and rich in tannins, essential oils andorganic acids. Serine/arginine rich splicing factors are essential inseqeunce specific splicing of pre-mRNAs. In this study we reportmolecular characterization of an serine/arginine rich SC35 splic-ing factor (OeSARsc35SF) that was isolated from a cDNAlibrary constructed from olive pedicels. Nucleotide BLAST andprotein BLAST (for comparison of the similarity of the candidategenes from other organisms) were conducted on NCBI web page.Amino acid composition analysis, nucleotide composition analy-sis, hydropathy analysis, open reading frame determiantion,through, molecular weight and the isoelectric points calculationswere conducted using online ExPasy software. Primer3 was usedto design forward and reverse primers to amplify the target genefrom different olive tissues at different times. Analysis with BioE-dit program revealed that A+T ratio was more than that of G+C.OeSARsc35SFwas a protein consisting of 267 amino acids. Asexpected, amino acid composition analysis revealed that serines and arginines were more than other amino acids. Kyte&Doolittlehyddropathy analysis revealed that the protein was hydrophilic.The molecular weight of the protein was 30 kDa with an isoelec-tric point (pI) of 11.5. The protein was found to have a signalpeptide. According to the Predotar analysis results, intracellularlocalization was found to be in the mitochondrial. The combinedresults suggest OeSARsc35SF might function as splicing factor asits homologs from the other plants. Confirming this hypothesiswith futher experimental characterization including biochemicalfunction analysis continues.Acknowledgements: This study was supported by TUB_ITAKwith grant number 110O108.Keywords:Olea europaeaL.,OeSARsc35SF, Alternative spl-cing, pre-mRNA splicing, BioEdit, Pedicel specific cDNAs.