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Abstract
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The human papillomavirus has the ability to infect the skin and the mucosal layer. Various types of the virus lead to wart lesions or cell transformations. In this regard, two types of HPV, Low-risk and high-risk has been introduced. Of HPV high-risk types, 16 and 18 are the main cause of cervical cancer, the second most common cancer in women worldwide. Despite a few prophylactic vaccines available, no therapy has been introduced for infected women yet. The aim of this study was to design and construct an expression vector for E7 protein of HPV-18 in yeast. This protein plays a key role in the development of cancer, and its successful expression could be promising to introduce a vaccine as a prevention and treatment of viral infection. For this purpose, the HPV-18 DNA was extracted from the patient's specimen and the E7- coding fragment was amplified using Nested-PCR in two steps. PCR product was double digested by NotI and XhoI and then cloned into PpiczαB vector. Cloning was confirmed by selection on zeosin, Colony PCR and restriction analysis. Plasmid DNA of colonies containing the E7 fragment was sequenced and compared with the sequences in the GenBank. The results indicate that the E7 gene of HPV-18 is successfully cloned in PpiczαB vector and the resulting plasmid can be used for expression studies in the yeast host.
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