Cloning and Construction of Eukaryotic Expression Vector of Buffalo NAAA Gene

In order to elucidate the function and molecular mechanisms of buffalo NAAA gene during folliculogenesis and embryogenesis, buffalo NAAA gene was studied with 3'-full Race, eukaryotic vector construction and cell transfection technology in this study. The results show that the coding region of buffalo NAAA is 1 071 bp in length, encoded 356 amino acids. The buffalo NAAA gene shares 98%, 96%, 95%, 87%, 87%, 85% and 84% similarity of nucleotide sequences with that of Bos taurus, Capra hircus, Ovis aries, Sus scrofa, Equus caballus, Felis catus and Homo sapiens, respectively. NAAA protein is weakly acidic, 26th~27th region was its signal peptide, located in the lysosome, and existed NAAA-beta and Ntn_AC_NAAA structural domains. The buffalo NAAA eukaryotic expression vector was successfully constructed. After transformed into 293T cell lines, NAAA-EGFP fusion protein was detectable. This study provided an important reference for surveying the regulation mechanism of NAAA gene during buffalo folliculogenesis and embryogenesis.