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...
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| Format: | Article |
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Mashhad University of Medical Sciences
2022-10-01
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| Series: | Majallah-i Zanān, Māmā̓ī va Nāzā̓ī-i Īrān |
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| Online Access: | https://ijogi.mums.ac.ir/article_21201_2642ffbce07b3f044639b53551ad249d.pdf |
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| author | Mohaddeseh Koohichapan Solmaz Moniri Javadhesari Farshad Darvishi Harzevili Catherine Madzak |
| author_facet | Mohaddeseh Koohichapan Solmaz Moniri Javadhesari Farshad Darvishi Harzevili Catherine Madzak |
| author_sort | Mohaddeseh Koohichapan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T16:58:14Z |
| format | Article |
| id | doaj.art-eea56243766c440f9dfb1c72a48ee9a3 |
| institution | Directory Open Access Journal |
| issn | 1680-2993 2008-2363 |
| language | fas |
| last_indexed | 2024-04-13T16:58:14Z |
| publishDate | 2022-10-01 |
| publisher | Mashhad University of Medical Sciences |
| record_format | Article |
| series | Majallah-i Zanān, Māmā̓ī va Nāzā̓ī-i Īrān |
| spelling | doaj.art-eea56243766c440f9dfb1c72a48ee9a32022-12-22T02:38:45ZfasMashhad University of Medical SciencesMajallah-i Zanān, Māmā̓ī va Nāzā̓ī-i Īrān1680-29932008-23632022-10-01258688010.22038/ijogi.2022.2120121201Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolyticaMohaddeseh Koohichapan0Solmaz Moniri Javadhesari1Farshad Darvishi Harzevili2Catherine Madzak3M.Sc. of Cellular and Molecular Biology, School of Basic Sciences, Azarbaijan Shahid Madani University, Tabriz, Iran.Assistant Professor, Department of Cellular and Molecular Biology, School of Basic Sciences, Azarbaijan Shahid Madani University, Tabriz, Iran.Professor, Department of Microbiology, School of Biological Sciences, Alzahra University, Tehran, Iran.INRAE France’s New National Research Institute for Agriculture, Food and Environment, Paris-Saclay University, Paris, France.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.https://ijogi.mums.ac.ir/article_21201_2642ffbce07b3f044639b53551ad249d.pdfrecombinant e6 proteinsurface display plasmidtype 18human papilloma viruscloning |
| spellingShingle | Mohaddeseh Koohichapan Solmaz Moniri Javadhesari Farshad Darvishi Harzevili Catherine Madzak Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica Majallah-i Zanān, Māmā̓ī va Nāzā̓ī-i Īrān recombinant e6 protein surface display plasmid type 18 human papilloma virus cloning |
| title | Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica |
| title_full | Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica |
| title_fullStr | Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica |
| title_full_unstemmed | Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica |
| title_short | Developing a plasmid for surface display containing the recombinant E6 protein of human papilloma virus type 18 for expression in yeast Yarrowia lipolytica |
| title_sort | developing a plasmid for surface display containing the recombinant e6 protein of human papilloma virus type 18 for expression in yeast yarrowia lipolytica |
| topic | recombinant e6 protein surface display plasmid type 18 human papilloma virus cloning |
| url | https://ijogi.mums.ac.ir/article_21201_2642ffbce07b3f044639b53551ad249d.pdf |
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