Cloning and expression of an anti-cancerous cytokine: human IL-29 gene in Chlamydomonas reinhardtii

Green algae, Chlamydomonas reinhardtii , with low cultivation cost, absence of endotoxins and insusceptibility to human pathogens is emerging as a potential system for the future production of recombinant proteins. The recent development of molecular tools enabling recombinant protein expression in...

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Bibliographic Details
Published in:AMB Express 2023-02, Vol.13 (1), p.23-23, Article 23
Main Authors: Akram, Maham, Khan, Mohsin Ahmad, Ahmed, Nadeem, Bhatti, Rashid, Pervaiz, Rabbia, Malik, Kausar, Tahir, Saad, Abbas, Rabia, Ashraf, Fareeha, Ali, Qurban
Format: Article
Language:English
Subjects:
Algae
Biological activity
Biomedical and Life Sciences
Biotechnology
Chlamydomonas reinhardtii
Chloroplast expression
Chloroplasts
Cloning
Endotoxins
Gel electrophoresis
Interleukin 29 (IL29)
Interleukins
Life Sciences
Low cost
Microbial Genetics and Genomics
Microbiology
Original
Original Article
Proteins
Sodium lauryl sulfate
Therapeutic protein
TN72 strain
Western blotting
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Summary:Green algae, Chlamydomonas reinhardtii , with low cultivation cost, absence of endotoxins and insusceptibility to human pathogens is emerging as a potential system for the future production of recombinant proteins. The recent development of molecular tools enabling recombinant protein expression in algae chloroplast has provided new research and advance opportunities for developing low-cost therapeutic proteins. In the present study, algae chloroplast expression system was evaluated for the recombinant production of an anti-cancerous therapeutic protein, Interleukin 29 ( IL29 ). The IL29 gene was cloned into algae chloroplast expression vector (pSRSapI). After the transformation, the positive clones were screened for homoplasmy and the presence of the IL29 gene by spot test and PCR analysis, respectively. The expressed SDS-PAGE and western blotting assay characterized IL-29 . The algae expressed IL-29 was biologically active in an anti-proliferating bioassay using HepG2 cells. The results suggest that the Chlamydomonas reinhardtii expression system is convenient, low-cost, eco-friendly, and safe to express IL29 . Key points The IL29 gene cloning in pSRSapI. IL-29 found as biologically active.
ISSN:2191-0855
2191-0855
DOI:10.1186/s13568-023-01530-1