MOESM1 of The peptide transporter 1a of the zebrafish Danio rerio, an emerging model in nutrigenomics and nutrition research: molecular characterization, functional properties, and expression analysis
2019-12-20T06:32:30Z (GMT) by
Additional file 1: Table S1. List of the specific primers used for cloning and qPCR analysis. Sequence accession numbers, primer sequences and amplicon sizes are shown. Figure S1. Nucleotide and predicted amino acid sequence of zebrafish pept1a (slc15a1a) obtained using ORFfinder (https://www.ncbi.nlm.nih.gov/orffinder/). Numbers on the left refer to the nucleotide (upper row) and amino acid (lower row) positions. Nucleotides are numbered, starting from the first ATG initiation codon. * indicates the stop codon. The specific primers used for cloning and PCR analyses (Additional file 1: Table S1) are indicated in red and green, respectively. In the amino acid sequence, putative transmembrane domains, obtained using the TMHMM v. 2.0 program as implemented in SMART, are indicated and named I to XII. Potential extracellular N-glycosylation sites (white boxes), potential cAMP/cGMP-dependent protein kinase phosphorylation sites at the cytoplasmic surface (light gray boxes) and potential protein kinase C phosphorylation sites at the cytoplasmic surface (dark gray boxes) were obtained using the ScanProsite tool. Figure S2. Current-voltage relationships of transport-associated currents in zebrafish PepT1a, in the presence of 3 mmol/L Gly-Gln in sodium (NaCl) saline buffer (black square) and tetramethylammonium (TMACl) saline buffer (empty circle) at pH 7.6 (see Methods for details). Values are means ± SEM from 5 oocytes from one batch each group. The transport-associated current values reported were obtained by subtracting the current recorded in the absence of the substrate to that recorded in its presence. Figure S3. Expression analysis by RT-PCR on pept1a (slc15a1a) mRNA in different sections of adult zebrafish intestine. a RT-PCR assay on cDNA templates from total RNA extracted from whole gut (gut), intestinal bulb (I. bulb), mid intestine (mid) and posterior intestine (posterior); a PCR product of ~ 350 bp related to pept1a (slc15a1a) mRNA is present in all intestinal samples; L: 1 Kb Plus DNA ladder (Thermo Fisher Scientific). b A graphic representation of the adult zebrafish gut anatomy with its major adjacent tracts.
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