مشخصات پژوهش

The human papillomavirus exclusively infects epithelial cells and leads to uncontrolled cell proliferation. HPV has more than 100 types that are divided into low risk and high risk types. The low risk HPV is the cause of warts and high risk HPV is associated with various cancers such as cervical cancer, vagina, skin, head and neck, anal and breast cancer. High risk HPV types 16 and 18 account for 70% of the cervical cancer, the second common cancer among women. Since early infections lead slowly to cancer, early detection and vaccination are very effective in reducing and preventing cell transformations. During viral contamination, E6 protein plays an important role in suppressing P53 protein and developing cell transformation. The purpose of this study was to construct an expression vector for E6 protein of HPV-18 to be expressed in Pichia pastoris. For this purpose, virus DNA was extracted from Pap smear sample of an infected woman. The E6 coding fragment was then amplified by two pairs of primers in a nested PCR. The obtained fragment was then cloned into the PpiczαB vector using NotI and XhoI restriction sites. The resulted clones were analyzed by quick check and the presence of E6 encoding fragment was confirmed by PCR and double digestion experiments. The sequence of cloned fragment was defined and recorded with the MK648315 accession number at GenBank. Our results indicated that E6 coding fragment is accurately cloned and the obtained vector can be used to express the E6 protein in Pichia pastoris.