en زیست شناسی ملکولی Molecular Biology <p style="text-align: justify;"><em><strong>Background:</strong></em> <em>Mycobacterium tuberculosis</em> is the causative agent of tuberculosis (TB). <em>Bacille Calmette-Guerin</em> (BCG) vaccine, is not effective in adults, therefore, many efforts have been made to produce an effective adult TB vaccine. The aim of this study was to develop a new tuberculosis DNA vaccine candidate encoding a recombinant HspX-PPE44-EsxV fusion antigen of <em>M. tuberculosis. </em></p> <p style="text-align: justify;"><em><strong>Methods:</strong></em> A fusion DNA segment consisting of HspX, linker, PPE44, linker, and EsxV, after codon optimization, was designed. The fusion DNA was cloned and its sequence confirmed. Then, expression of a recombinant pcDNA3.1 (+)/HspX-PPE44-EsxV plasmid in Chinese hamster ovary (CHO) cells was verified by RT-PCR and Western-blot analysis.</p> <p style="text-align: justify;"><em><strong>Results:</strong></em> A 1968 bp band in RT-PCR and a 68 kDa band on Western-blot analysis confirmed transcription and expression of recombinant <em>hspX-ppe44-esxV</em> in eukaryotic cells.</p> <p style="text-align: justify;"><em><strong>Conclusions: </strong></em>A recombinant DNA segment encoding the HspX-PPE44-EsxV fusion antigen of <em>M. tuberculosis</em> was constructed and considered to be tested as a new TB DNA vaccine candidate.</p> DNA vaccine, EsX, Hsp, Mycobacterium tuberculosis, PPE44 89 97 http://rbmb.net/browse.php?a_code=A-10-1-55&slc_lang=en&sid=1 Bagher Moradi 10031947532846006137 10031947532846006137 No Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Mojtaba Sankian 10031947532846006138 10031947532846006138 No Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Yousef Amini 10031947532846006139 10031947532846006139 No Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Zahra Meshkat meshkatz@mums.ac.ir 10031947532846006140 10031947532846006140 Yes Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran