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Updated: May 26, 2025
CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
Published on: June 25, 2017
Designing and cloning of fusion protein CpsA-CpsC-L-ACAN
Elmira Babakanrad1, Taher Mohammadian1, Davoud Esmaeili2
1Department of Microbiology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.
Background:
Cervical cancer is the fourth most common cause of cancer and the fourth most common cause of cancer deaths in women. Some reports have shown the effect of Streptococcus agalactiae proteins and capsule products against cancer cell lines.
Methods:
This study aimed to design and produce a fusion of recombinant protein (containing the capsules of Streptococcus agalactiaes with a linker and Antti cancer sequences) CpsA-CpsC-L-ACAN in the plasmid pET-22b (+) vector. Construct pET-22b (+) was designed by researchers, optimized with bioinformatics software, and synthesized by a Biometrics company. For the confirmation of recombinant protein, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting were performed. Antibacterial activity was performed according to protocol CLSI2020. Based on bioinformatics analysis, the recombinant protein had good spatial structure and stability and half-life. Iterative Threading ASSEmbly Refinement (I-TASSER) results predicted a surfing topology with C-score values (-3.60).
Results:
The results of Gastro-Oesophageal Reflux 4 (GOR4) analysis showed a little extended strand (30.63%), a random coil with a percentage (50.18%), and an alpha helix with a percentage (19.19%). The recombinant protein was confirmed by SDS-PAGE and western blotting with Anti-his tag.
Conclusion:
The purpose of peptide fusion design in this article is to help in the development of anti-cervical cancer medicine. Of course, this issue needs to be investigated in animal and human phases. In this article, only bioinformatics investigations and validation of the results in the laboratory have been discussed.
