Molecular cloning, sequence characterization, and tissue expression analysis of three water buffalo (Bubalus bubalis) genes – ST6GAL1, ST8SIA4, and SLC35C1

Abstract. Recent studies have shown that ST6 beta-galactosamide alpha-2,6-sialyltransferase 1 (ST6GAL1), ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyltransferase 4 (ST8SIA4), and solute carrier family 35, member C1 (SLC35C1) play essential roles in the metabolism of milk glycoconjugates in mammals. However, studies on their coding genes in water buffalo have not been reported. In the present study, cloning and sequencing showed that the coding sequences (CDSs) of buffalo ST6GAL1, ST8SIA4, and SLC35C1 were 1218, 1080, and 1095 bp in length, which encoded a precursor protein composed of 405, 359, and 364 amino acids, respectively. The deduced sequences of these three proteins in turn showed 97.6–98.5, 98.6–99.7, and 97.8–99.2 % similarities with other bovine species. Both buffalo ST6GAL1 and ST8SIA4 were predicted to be a member of glycosyltransferase family 29 and were all hydrophilicity proteins functioning in the Golgi apparatus. Buffalo SLC35C1 was a hydrophobic membrane protein located in the Golgi membrane, containing a TPT domain that is found in a number of sugar phosphate transporters. In addition, semi-quantitative RT-PCR analysis in 13 lactating buffalo tissues revealed that the ST6GAL1 and ST8SIA4 were expressed in 9 tissues, while SLC35C1 was expressed in 11 tissues. The expression levels of these three genes in the mammary gland were significantly higher in lactating than in non-lactating stage. Collectively, our data indicate that ST6GAL1, ST8SIA4, and SLC35C1 are potentially involved in the process of buffalo lactation.