Cloning, analysis and expression detection of the full-length cDNA of <I>Sox9</I> from the Siberian sturgeon (<I>Acipenser baerii</I> Brandt)

Abstract

In order to further research the role of Sox9 in the development of the Siberian sturgeon (Acipenser baerii Brandt)’s lateral line neuromast,the full-length Sox9 cDNA of Siberian sturgeon had been successfully cloned by RT-PCR and rapid amplification of cDNA ends (RACE) methods. The full-length Sox9 cDNA of Siberian sturgeon had 1 409 bp,which consisted of a 1 083 bp open reading frame (ORF) encoding 360 amino acids,a 19 bp 5′untranslated region (5′UTR) and a 307 bp 3′untranslated region (3′UTR). The molecular weight of this mature peptide was estimated to be 41 221. 7U. Neighbor-Joining (NJ) method had been used for multiple alignment analyses,and Siberian sturgeon was closest to Amur sturgeon ( Acipenser schrenckii). The Siberian sturgeon’s Sox9 gene was highly conservative to other species. These results were much similar to other studies that had already been done in blue tilapia (Oreochromis aurea),yellow catfish (Pelteobagrus fulvidraco) and swamp eel (Moncpterus albus). Meanwhile,in order to study the connection between the feature of the expression of the Sox9 to the development of the Siberian sturgeon’s lateral line neuromast,the fluorescent quantitative RT-PCR analysis was used for detecting Sox9 relative expression levels in the different tissues and different embryonic development stages. Sox9 had relatively low expression levels in liver,pancreas,kidney and muscle. However,in the brain,Sox9 had very high expression levels,and the relative expression levels of the brain reached 8. 9 times the quantity of the relative expression in the pancreas and 21 times the quantity of the relative expression in the kidney. In the different stages of the Siberian sturgeon embryonic development,Sox9 expression had different relative expression levels. Its relative expression had no major differences and reached relatively high in gastrula stage,neurula stage and formation of visual bubble. Especially in the gastrula stage and neurula stage,Sox9 relative expression reached 1. 2 times and 1. 4 times the quantity of formation of heart. All of these results were nearly same as the researches of zebrafish ( Danio rerio),chicken ( Gallus gallus) and mouse ( Mus musculus). At the same time,the expression features showed Sox9 might play a very important role in the development of the neuromast of the Siberian sturgeon which had already been found in the development processes of the zebrafish neuromast. This research will lay a foundation for the future study on the role of Siberian sturgeon’s Sox9 gene in the differentiation of lateral line ganglion development,cells migration processes and so on. At the same time,the study on the role of the Sox9 in the cell signaling pathway of the development of the Siberian sturgeon’s lateral line neuromast is required too.