Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica
Introduction: Human papillomaviruses (HPV) with more than 100 types are categorized as low-risk and high-risk types. Types 16 and 18 of the virus alone are involved in 70% of cervical cancers. Currently, development of recombinant proteins of HPV for vaccination or therapy purposes has attracted sci...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | fas |
| Published: |
Mashhad University of Medical Sciences
2022-10-01
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| Series: | Majallah-i Zanān, Māmā̓ī va Nāzā̓ī-i Īrān |
| Subjects: | |
| Online Access: | https://ijogi.mums.ac.ir/article_21201_2642ffbce07b3f044639b53551ad249d.pdf |
| Summary: | Introduction: Human papillomaviruses (HPV) with more than 100 types are categorized as low-risk and high-risk types. Types 16 and 18 of the virus alone are involved in 70% of cervical cancers. Currently, development of recombinant proteins of HPV for vaccination or therapy purposes has attracted scientists. Therefore, the present study was performed aimed to construct a surface display plasmid encoding E6 protein of human papillomavirus type 18.Methods: The DNA fragment encoding E6 protein of HPV18 was amplified by nested-PCR using DNA of a HPV18 positive person as PCR template. Then, the amplified fragment was double digested with HindIII and SfiI and cloned into the surface display plasmid pINA1317-YLCWP110.Results: Cloning of E6 protein encoding gene fragment into pINA1317-YLCWP110 plasmid was confirmed using PCR and restriction endonuclease double digestion. Also, the results of Sanger sequencing of the recombinant pINA1317-YLCWP110-E6 plasmid and alignment to gene bank further ensured the sequence accuracy, cloning position and reading frame of the gene in the recombinant vector.Conclusion: DNA fragment encoding E6 protein of HPV18 was successfully cloned into surface display plasmid pINA1317-YLCWP110 in appropriate locus and frame. Altogether, the recombinant pINA1317-YLCWP110-E6 plasmid constructed in this study can be expressed in the yeast host Yarrowia lipolytica and the resulted E6 protein may be used as a prophylactic or therapeutic vaccine or molecular marker. |
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| ISSN: | 1680-2993 2008-2363 |