Posters
Presentation Type
Poster
Discipline Track
Clinical Science
Abstract Type
Research/Clinical
Abstract
Background: Hauzen et al., went against the dogma that cancer only develops by genetic factors and postulated oncogenic human papillomavirus (HPV) as the etiological agent of cervical cancer (2008). The eleven new human polyomaviruses (HPyVs) identified have been shown to infect humans subclinically from an early age. The small t-antigen (tAg) proteins of polyomaviruses control cellular phosphorylation mechanisms leading to uncontrolled replication cycles and cancer. Mechanisms of transformation and progression to tumor cells, and cellular tropism of the new HPyVs, remain unknown. Availability of recombinant tAg (rtAg) can contribute to understanding them.
Methods: tAg coding sequences derived from Merkel cell polyomavirus (MCPyV) genome were codon-optimized, synthesized, cloned in an expression vector (pGEX), and transformed into E. coli. Clones were fermented, induced for expression of the tAG cassette, and purified by IMAC (Immobilized metal affinity chromatography).
Results: rtAg produced in bacteria from different expression strains demonstrated distinct expression levels. The rtAg of ~20KDa was produced at the level of 11.3 mg L-1 and folded correctly since antibody 5 anti-MCPyV recognized rtAg.
Conclusions: MCPyV tAg was efficiently expressed in E. coli. The availability of rtAg from MCPyV will be useful in immunity diagnostics, structure studies, and investigation of metabolic pathways interference and cell tropism features of HPyVs infections. This especially until cell culture systems for new HPyVs are developed.
Acknowledgements: Our sincere gratitude to the Dana-Farber Cancer Institute for gene and antibody samples as well as the National Council of Science and Technology (CONACYT) for the scholarship 304814 to LMRM.
Academic/Professional Position
Post-doc
Recommended Citation
Rodriguez, Luis Mario; Barrera-Saldaña, Hugo A.; and DeCaprio, James A., "Production of codon optimized Polyomavirus small t antigen in Escherichia coli." (2023). Research Symposium. 81.
https://scholarworks.utrgv.edu/somrs/theme1/posters/81
Included in
Production of codon optimized Polyomavirus small t antigen in Escherichia coli.
Background: Hauzen et al., went against the dogma that cancer only develops by genetic factors and postulated oncogenic human papillomavirus (HPV) as the etiological agent of cervical cancer (2008). The eleven new human polyomaviruses (HPyVs) identified have been shown to infect humans subclinically from an early age. The small t-antigen (tAg) proteins of polyomaviruses control cellular phosphorylation mechanisms leading to uncontrolled replication cycles and cancer. Mechanisms of transformation and progression to tumor cells, and cellular tropism of the new HPyVs, remain unknown. Availability of recombinant tAg (rtAg) can contribute to understanding them.
Methods: tAg coding sequences derived from Merkel cell polyomavirus (MCPyV) genome were codon-optimized, synthesized, cloned in an expression vector (pGEX), and transformed into E. coli. Clones were fermented, induced for expression of the tAG cassette, and purified by IMAC (Immobilized metal affinity chromatography).
Results: rtAg produced in bacteria from different expression strains demonstrated distinct expression levels. The rtAg of ~20KDa was produced at the level of 11.3 mg L-1 and folded correctly since antibody 5 anti-MCPyV recognized rtAg.
Conclusions: MCPyV tAg was efficiently expressed in E. coli. The availability of rtAg from MCPyV will be useful in immunity diagnostics, structure studies, and investigation of metabolic pathways interference and cell tropism features of HPyVs infections. This especially until cell culture systems for new HPyVs are developed.
Acknowledgements: Our sincere gratitude to the Dana-Farber Cancer Institute for gene and antibody samples as well as the National Council of Science and Technology (CONACYT) for the scholarship 304814 to LMRM.