A general and effective strategy to identify cDNAencoding for mitochondrial silver eel citrate carrier(CIC) has been developed. In particular using primersdirected towards the highly conserved signaturemotif of the ortholog citrate cDNA sequences, thefull-length silver eel cDNA from liver was obtained.This is 1193 bp in length with 5’ and 3’ untranslatedregions of 90 and 149 bp, respectively. The open readingframe encodes a mature protein of 297 aminoacids, preceded by a presequence of 20 amino acids.Additionally the mature CIC overexpressed in Escherichiacoli and reconstituted into phospholipidsvesicles showed the same substrate specificity of thenative protein previously characterized in silver eel.The tissue distribution of silver eel CIC mRNAs wasinvestigated and transcripts were detected at highlevels in swim bladder whereas a weaker signal wasfound in brain, gill, intestine and liver.
A general and effective strategy to identify cDNAencoding for mitochondrial silver eel citrate carrier(CIC) has been developed. In particular using primersdirected towards the highly conserved signaturemotif of the ortholog citrate cDNA sequences, thefull-length silver eel cDNA from liver was obtained.This is 1193 bp in length with 5’ and 3’ untranslatedregions of 90 and 149 bp, respectively. The open readingframe encodes a mature protein of 297 aminoacids, preceded by a presequence of 20 amino acids.Additionally the mature CIC overexpressed in Escherichiacoli and reconstituted into phospholipidsvesicles showed the same substrate specificity of thenative protein previously characterized in silver eel.The tissue distribution of silver eel CIC mRNAs wasinvestigated and transcripts were detected at highlevels in swim bladder whereas a weaker signal wasfound in brain, gill, intestine and liver.
An effective strategy for cloning the mitochondrial citrate carrier: identification, characterization and tissue distribution in silver eel
MADEO, Marianna;CAPPELLO, Anna Rita;DOLCE, Vincenza
2011-01-01
Abstract
A general and effective strategy to identify cDNAencoding for mitochondrial silver eel citrate carrier(CIC) has been developed. In particular using primersdirected towards the highly conserved signaturemotif of the ortholog citrate cDNA sequences, thefull-length silver eel cDNA from liver was obtained.This is 1193 bp in length with 5’ and 3’ untranslatedregions of 90 and 149 bp, respectively. The open readingframe encodes a mature protein of 297 aminoacids, preceded by a presequence of 20 amino acids.Additionally the mature CIC overexpressed in Escherichiacoli and reconstituted into phospholipidsvesicles showed the same substrate specificity of thenative protein previously characterized in silver eel.The tissue distribution of silver eel CIC mRNAs wasinvestigated and transcripts were detected at highlevels in swim bladder whereas a weaker signal wasfound in brain, gill, intestine and liver.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
