Volume 16, Issue 62 (5-2026)
NCMBJ 2026, 16(62): 63-73 |
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Ezzatizadeh F, Bolhassani A, Sotoodehnejadnematalahi F, Fateh A. Construction of an Expression Vector Carrying the Human Papillomavirus Type 16 Oncogene E5 and Evaluation of Its Expression in an E. coli Expression System. NCMBJ 2026; 16 (62) :63-73
URL: http://ncmbjpiau.ir/article-1-1822-en.html
URL: http://ncmbjpiau.ir/article-1-1822-en.html
Department of Hepatitis, AIDS and Blood-borne Diseases, Pasteur Institute of Iran, Tehran, Iran
Abstract: (12 Views)
Background and Aim: Human papillomavirus type 16 (HPV16) oncoproteins play a pivotal role not only in the severity of HPV-related diseases but also in the early diagnosis of virus-induced cancers. This study aimed to clone the HPV16 E5 oncogene into a prokaryotic expression vector and assess its expression in an Escherichia coli (E. coli) system, with the future goal of facilitating its purification and functional characterization.
Materials and Methods: In this study, the HPV16 E5 gene was cloned into the pET28a prokaryotic expression vector (plasmid expression T7). Then, expression of the recombinant E5 protein was analyzed in the E. coli expression system using SDS-PAGE and western blotting techniques.
Results: Our results showed that the E5 gene, which is approximately 240 base pairs in size and was cloned into the pET28a(+) vector, was expressed in E. coli BL21 under the following conditions: 37º C, 0.5 mM IPTG, and 18 hours post-induction. SDS-PAGE and western blot analysis confirmed a distinct protein band at approximately 10 kDa, consistent with the expected size of the recombinant E5 protein.
Conclusion: The successful expression of the hydrophobic E5 oncoprotein in a bacterial system, despite the absence of post-translational modifications such as phosphorylation and glycosylation, suggests the purification of this protein for evaluation of its function in vaccine design in the future.
Materials and Methods: In this study, the HPV16 E5 gene was cloned into the pET28a prokaryotic expression vector (plasmid expression T7). Then, expression of the recombinant E5 protein was analyzed in the E. coli expression system using SDS-PAGE and western blotting techniques.
Results: Our results showed that the E5 gene, which is approximately 240 base pairs in size and was cloned into the pET28a(+) vector, was expressed in E. coli BL21 under the following conditions: 37º C, 0.5 mM IPTG, and 18 hours post-induction. SDS-PAGE and western blot analysis confirmed a distinct protein band at approximately 10 kDa, consistent with the expected size of the recombinant E5 protein.
Conclusion: The successful expression of the hydrophobic E5 oncoprotein in a bacterial system, despite the absence of post-translational modifications such as phosphorylation and glycosylation, suggests the purification of this protein for evaluation of its function in vaccine design in the future.
Keywords: HPV16, E5 oncoprotein, Cloning, Recombinant protein expression, Escherichia coli expression system
Type of Study: Research Article |
Subject:
Cellular and molecular
Received: 2026/06/21 | Accepted: 2026/05/31 | Published: 2026/05/31
Received: 2026/06/21 | Accepted: 2026/05/31 | Published: 2026/05/31
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