Introducing Single Nucleotide variations into the Vitamin C Transporter SLC23A

Abstract

The solute carrier SLC23A1, encoded by the SLC23A1 gene, is the key membrane transporter mediating the reabsorption of ascorbic acid, the reduced form of vitamin C, in the renal proximal tubule. It had been shown that a single nucleotide polymorphism (SNP) in SLC23A1 could alter the proteins ability to transport ascorbic acid. Recently, through various human re-sequencing projects, many novel single nucleotide variations (SNV) have been discovered and their information has been deposited in genomic databases. However, their impact on the proteins functions remains to be determined. In a first step for functional studies, SNV need to be introduced into an existing SLC23A1 expression plasmids. The objective of this study was to introduce selected SNVs into the coding region, or Open Reading Frame (ORF), of the SLC23A1 expression plasmid pReceiver-M55-SLC23A1. Six SNVs located proximal to the 5’ and 3’ ends of the SLC23A1 ORF were successfully introduced.