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Volume 10
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10.3390/microorganisms10020361
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Article

Transformation of Dunaliella salina by Agrobacterium tumefaciens for the Expression of the Hemagglutinin of Avian Influenza Virus H5

by
Inkar Castellanos-Huerta
1,*,
Gabriela Gómez-Verduzco
2,
Guillermo Tellez-Isaias
3,
Guadalupe Ayora-Talavera
4,
Bernardo Bañuelos-Hernández
5,
Víctor Manuel Petrone-García
6,
Gilberto Velázquez-Juárez
7 and
Isidro Fernández-Siurob
8
1
Programa de Maestría y Doctorado en Ciencias de la Producción y de la Salud Animal, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de Mexico 04510, Mexico
2
Departamento de Medicina y Zootecnia de Aves, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Avenida Universidad 3000, Ciudad de Mexico 04510, Mexico
3
Department of Poultry Science, University of Arkansas, Fayetteville, AK 72701, USA
4
Centro de Investigaciones Regionales, Dr. Hideyo Noguchi, Universidad Autonoma de Yucatán (UADY), Mérida 97000, Mexico
5
Escuela de Veterinaria, Universidad De La Salle Bajío, Avenida Universidad 602, Lomas del Campestre, León 37150, Mexico
6
Departamento de Ciencias Pecuarias, Facultad de Estudios Superiores Cuautitlán UNAM, Cuautitlán Izcalli 54714, Mexico
7
Departamento de Química, Centro Universitario de Ciencias Exactas e Ingenierías, Universidad de Guadalajara, Blvd. Marcelino García Barragán #1421, Guadalajara 44430, Mexico
8
Viren SA de CV, Presidente Benito Juárez 110B, José María Arteaga, Querétaro 76135, Mexico
*
Author to whom correspondence should be addressed.
Microorganisms 2022, 10(2), 361; https://doi.org/10.3390/microorganisms10020361
Submission received: 19 December 2021 / Revised: 4 January 2022 / Accepted: 31 January 2022 / Published: 4 February 2022
(This article belongs to the Section Veterinary Microbiology)
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Abstract

Avian influenza (AI) is one of the main threats to the poultry industry worldwide. Vaccination efforts are based on inactivated, live attenuated, and recombinant vaccines, where the virus hemagglutinin (HA) is the main component of any vaccine formulation. This study uses Dunaliella salina to express the AIV HA protein of an H5 virus. D. salina offers a system of feasible culture properties, generally recognized as safe for humans (GRAS), with N-glycosylation and nuclear transformation by Agrobacterium tumefaciens. The cloning and transformation of D. salina cells with the H5HA gene was confirmed by polymerase chain reaction (PCR). SDS-PAGE and Western blot confirmed HA5r protein expression, and the correct expression and biological activity of the HA5r protein were confirmed by a hemagglutination assay (HA). This study proves the feasibility of using a different biological system for expressing complex antigens from viruses. These findings suggest that a complex protein such as HA5r from AIV (H5N2) can be successfully expressed in D. salina.
Keywords: avian influenza; Dunaliella salina; Agrobacterium tumefaciens; hemagglutinin; recombinant avian influenza; Dunaliella salina; Agrobacterium tumefaciens; hemagglutinin; recombinant

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MDPI and ACS Style

Castellanos-Huerta, I.; Gómez-Verduzco, G.; Tellez-Isaias, G.; Ayora-Talavera, G.; Bañuelos-Hernández, B.; Petrone-García, V.M.; Velázquez-Juárez, G.; Fernández-Siurob, I. Transformation of Dunaliella salina by Agrobacterium tumefaciens for the Expression of the Hemagglutinin of Avian Influenza Virus H5. Microorganisms 2022, 10, 361. https://doi.org/10.3390/microorganisms10020361

AMA Style

Castellanos-Huerta I, Gómez-Verduzco G, Tellez-Isaias G, Ayora-Talavera G, Bañuelos-Hernández B, Petrone-García VM, Velázquez-Juárez G, Fernández-Siurob I. Transformation of Dunaliella salina by Agrobacterium tumefaciens for the Expression of the Hemagglutinin of Avian Influenza Virus H5. Microorganisms. 2022; 10(2):361. https://doi.org/10.3390/microorganisms10020361

Chicago/Turabian Style

Castellanos-Huerta, Inkar, Gabriela Gómez-Verduzco, Guillermo Tellez-Isaias, Guadalupe Ayora-Talavera, Bernardo Bañuelos-Hernández, Víctor Manuel Petrone-García, Gilberto Velázquez-Juárez, and Isidro Fernández-Siurob. 2022. "Transformation of Dunaliella salina by Agrobacterium tumefaciens for the Expression of the Hemagglutinin of Avian Influenza Virus H5" Microorganisms 10, no. 2: 361. https://doi.org/10.3390/microorganisms10020361

APA Style

Castellanos-Huerta, I., Gómez-Verduzco, G., Tellez-Isaias, G., Ayora-Talavera, G., Bañuelos-Hernández, B., Petrone-García, V. M., Velázquez-Juárez, G., & Fernández-Siurob, I. (2022). Transformation of Dunaliella salina by Agrobacterium tumefaciens for the Expression of the Hemagglutinin of Avian Influenza Virus H5. Microorganisms, 10(2), 361. https://doi.org/10.3390/microorganisms10020361

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