TY - JOUR AU - Cuong, Vo Viet AU - Hue, Le Thi AU - Huyen, Do Thi AU - Giang, Le Quynh AU - Quy, Nguyen Thi AU - Hai, Truong Nam PY - 2013/07/09 Y2 - 2024/03/29 TI - Expression of codon optimized HA5.1 gene with biofunctional activity in Pichia pastoris X33 JF - Academia Journal of Biology JA - AJB VL - 35 IS - 2 SE - Articles DO - 10.15625/0866-7160/v35n2.3113 UR - https://vjs.ac.vn/index.php/vjbio/article/view/3113 SP - 255-264 AB - <p>Exotic gene expression in <em>P. pastoris</em> is dependent on multistep processes involving regulation at the level of transcription, protein translation, and posttranslational modifications. In order to further improve the expression level of HA recombinant protein from A/H5N1 avian virus we optimized the HA5.1 protein coding sequence and expressed it in <em>P. pastoris</em> X33. After <em>optimization</em>, <em>Codon</em> Adaptation Index (CAI) value was improved from 0.69 to 0.98, without modifying the amino acid sequence of the encoded protein. The synthetic <em>mha</em>5.1 with a length of 1 kb was inserted in to pPICZαmha5.1 and cloned in <em>E. coli</em> DH10b then integrated into <em>P. pastoris</em> X33. HA5.1 recombinant protein of 50-70 kDa was synthesized at 30<sup>o</sup>C under induction of 1% methanol during 72 hours. Recombinant protein had biological function to agglutinate chicken’s blood cells at maximum 2<sup>6</sup> dilutions and had antigenicity to bind with antibody against HA5 of virus H5N1. These important results are the basis for using recombinant HA5.1 protein as a subunit vaccine.</p> ER -