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Coordinación General de Investigación de la Facultad de Salud Pública y Nutrición y la Dirección General de Sistemas e Informática de la Universidad Autónoma de Nuevo León

2005, Number 3

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Rev Salud Publica Nutr 2005; 6 (3)

Expression of HIV type 1 glycoprotein 120 from a mexican AIDS patient in a baculovirus system

Ramos-Alfano G, Tamez-Guerra P, Rivera-Morales L, Tamez-Guerra R, Gomez-Flores R, Rodriguez-Padilla C

Language: English
References: 43
Page:
PDF size: 297.60 Kb.


ABSTRACT

The aim of the present study was the implementation of an insect cell-baculovirus expression vector system (IC-BEVS) for the expression of the human immunodeficiency virus (HIV) type 1 gp120 recombinant protein. For this, we isolated the DNA sequences from the gp120 protein from a Mexican AIDS patient, and from the reference plasmid HXB2, which were independently cloned in the plasmid pFastBac1. We used the plasmids to transform E. coli DH10Bac containing the AcMNPV baculovirus bMON14272 bacmid, and then induced a site-directed transposition to transfer the HIV-gp120 genome to the baculovirus´s bacmid. Baculovirus recombinant was detected using PCR analysis and colony phenotyping, with both the Bac-69-1 and the reference plasmid Bac-HX-4. The baculoviral naked genome was then encapsulated within liposomes to transfect the insect cell culture Sf9. Western blot analysis, developed by chemiluminescence and hydrogen peroxide hydrolysis was then utilized to identify the recombinant gp120 expression. Results demonstrated the generation of two recombinant baculoviruses expressing either gp120 from a Mexican AIDS patient or as reference plasmid. The potential of using the gp120 recombinant baculovirus as HIV vaccine in Mexico is discussed.


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