TY - JOUR T1 - Design of Eukaryotic Vector Harboring L1-E7 Polyepitope Fusion Construct of High Risk Human Papillomaviruses TT - طراحی و تهیه وکتور یوکاریوتی حامل ساختار فیوژن پلی اپی توپی L1-E7 ویروسهای پاپیلومای انسانی پر خطر JF - NCMBJ JO - NCMBJ VL - 11 IS - 44 UR - http://ncmbjpiau.ir/article-1-1414-en.html Y1 - 2021 SP - 95 EP - 106 KW - Human papillomavirus KW - L1 protein KW - E7 protein KW - Eukaryotic expression vector KW - Iau Science. N2 - Abstract: Aim and Background: Human Papillomaviruses (HPV) infection was known to be the leading cause of cervical cancer. Cervical cancer is the fourth most common cancer among women in the world. Designing a DNA vaccine with therapeutic properties as well as prevention can have a significant impact on reducing morbidity and mortality. Materials and Methods: Gene sequences including the immunogenic and conserved epitopes of L1 and E7 proteins of high risk papillomaviruses were designed. After synthesis of the L1-E7 fusion construct in pUC57 cloning vector, its subcloning was performed in pcDNA3.1 (-) eukaryotic vector. The concentration and purity of the recombinant pcDNA-L1E7 plasmid was determined by NanoDrop spectrophotometry. Results: The gene sequence after cutting from pUC57-L1E7 by BamHI/HindIII restriction enzymes, was subcloned in the same cloning site of pcDNA3.1 (-) vector. The presence of gene was confirmed by digestion with BamHI/HindIII enzymes, as a 639 bp fragment on 1% agarose gel. The recombinant pcDNA-L1E7 plasmid was purified by endotoxin-free extraction kit for isolation of bacterial lipopolysaccharide. The concentration of the purified DNA was obtained about 1474 ng/ µl. Conclusion: Cloning of the L1-E7 fusion construct in the eukaryotic vector was successful. In the next steps, the recombinant vector will be used for gene vaccine studies in vivo. M3 ER -