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All Journal Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences THE JOURNAL OF MUHAMMADIYAH MEDICAL LABORATORY TECHNOLOGIST Meditory : The Journal of Medical Laboratory Klinikal Sains (Jurnal Analis Kesehatan) Jurnal Ilmu Kedokteran dan Kesehatan Jambura Journal of Health Sciences and Research Jurnal Medika Malahayati Husada Mahakam: Jurnal Kesehatan JURNAL ANALIS LABORATORIUM MEDIK Borneo Journal of Medical Laboratory Technology Medicra (Journal of Medical Laboratory Science/Technology) Health Care : Jurnal Kesehatan Jurnal Kesehatan dan Pembangunan Jurnal Kesehatan Tambusai Jurnal Sains dan Kesehatan Borneo Journal of Science and Mathematics Education Journal of Sustainability Science and Technology Gema Kesehatan
Kesuma, Suryanata
Poltekkes Kemenkes Kalimantan Timur
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Dentistry Education Energy Environmental Science Health Professions Immunology & microbiology Library & Information Science Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Nursing Physics Public Health

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Design, Construction and Expression of Spike Highly Conserved Region (HCR) SARS-CoV-2 and Cholera Toxin Subunit B Fusion Protein in Lactococcus lactis NZ3900: Construction of recombinant plasmid in Lactococcus lactis Kesuma, Suryanata; Adianingsih, Oktavia Rahayu; Winarsih, Sri; Widodo, Nashi; Yurina, Valentina
Journal of Tropical Life Science Vol. 15 No. 2 (2025): In Press
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/2xf1qp96

Abstract

Prevention of SARS-CoV-2 transmission has primarily been achieved through vaccination, which is generally administered via injection and may cause discomfort. No commercially available SARS-CoV-2 vaccines can be administered via the mucosal route. However, recent advancements have demonstrated that vaccination with Lactococcus lactis enables vaccine delivery through the mucosa. A promising target for SARS-CoV-2 vaccine materials is the highly conserved region (HCR) of the SARS-CoV-2 spike (SARS-CoV-2 HCR Spike). Vaccine efficacy is enhanced by adding Cholera Toxin Subunit B (CTB) as an adjuvant. HCR and CTB proteins were recombinantly fused using a synthetic gene with optimized codons. This study aimed to construct a fusion protein of the SARS-CoV-2 spike protein and CTB in L. lactis strain NZ3900. The construction and expression of fusion proteins were analyzed using sequencing and protein electrophoresis. Codon optimization resulted in a Codon Adaptation Index value of 0.93 and a GC content of 27.06%. The cloning results revealed the formation of L. lactis colonies expressing the Fusion protein of the SARS-CoV-2 HCR Spike and CTB, which formed yellow colonies on the selection Elicker medium. PCR and sequencing confirmed the presence of the hcr-ctb gene, with a length of 981 bp and 100% sequence similarity. The Fusion protein of the SARS-CoV-2 HCR Spike and CTB was successfully expressed with a molecular weight of >35 kDa. In conclusion, we successfully constructed a Fusion protein of the SARS-CoV-2 HCR Spike and CTB in L. lactis NZ3900 as a potential vaccine candidate for oral administration to prevent SARS-CoV-2 infection.
Co-Authors Adi Firmansyah Agus Rudi Hartono Amalia Diah Suci Rahmah Anggrieni, Nurul Armiah Gita Anggraini DESI WAHYUNI Didi Irwadi Dwi Wulan Sari Eka Farpina, Eka Farpina Elita Octavia Farhan Baehaki Farpina, Eka Fera, Fera Hartono, Agus Rudi I Gede Andika Sukarya Irawiraman, Hadi Lamri Lamri Lamri Lamri Liah Kodariah Mahdiah Syumarliyanty Marezha Aulia Putri Sugiarsono Maulida Maulida Julia Saputri Nanda Nur Adha Sari Febriyanti Nashi Widodo Nia Ardelia Nursalinda Kusumawati Nurul Alidasyah Nurul Hikmah Oktavia Rahayu Adianingsih Patricia Aleksandra Prihandono, Dwi Setiyo Rahmad Rezaldy Sari, Putri Sekar Silvi Marshela Siti Rodiyah, Siti Sresta Azahra Sri Winarsih Sultan Sultan Suparno Putera Makkadafi Suparno Putera Makkadafi Supri Hartini Tatang Abdullah VALENTINA YURINA Widyadari, Effie Raissa