Identification and characterization of amphibian SLC26A5 using RNA-Seq

We mapped and sequenced the SLC26A5 of the American bullfrog from its inner ear cDNA using RNA-Seq. The frog SLC26A5 cDNA was 2,292 bp long, encoding a polypeptide of 763 amino acid residues, with 40% identity to mammals. After isolating the prestin gene of the frog, we generated a stable cell line transfected with this new coding gene and found it possessing similar electrophysiological features as the hair cells from the frog's auditory organ. Our experiment demonstrated that the new coding gene could encode a functionally active protein conferring NLC to both frog HCs and the mammalian cell line. Overall design: The SLC26A5 gene of bullfrog has been mapped, sequenced and cloned successively using RNA-Seq. We measured the nonlinear capacitance (NLC) of prestin both in the hair cells of frog's inner ear and HEK293T cells transfected with this new coding gene.本研究通过RNA测序（RNA-Seq）技术，从美洲牛蛙的内耳cDNA中完成了溶质载体家族26成员A5（SLC26A5）的定位与测序。该牛蛙SLC26A5的cDNA序列全长为2292 bp，编码一条含763个氨基酸残基的多肽，与哺乳动物同源序列的相似度为40%。 在分离得到牛蛙的普雷斯汀蛋白（prestin）基因后，我们构建了稳定转染该编码基因的细胞系，发现其电生理特性与牛蛙听觉器官的毛细胞相似。 本实验证实，该新型编码基因可编码具有功能活性的蛋白，能够赋予牛蛙毛细胞（HCs）以及转染该基因的哺乳动物细胞系非线性电容（NLC）特性。 实验整体设计如下：本研究通过RNA-Seq技术依次完成了牛蛙SLC26A5基因的定位、测序与克隆；并分别在牛蛙内耳毛细胞以及转染该编码基因的人胚肾293T（HEK293T）细胞中，检测了普雷斯汀蛋白（prestin）的非线性电容（NLC）特性。