EXPRESSION OF HIV TYPE 1 GLYCOPROTEIN 120 FROM A MEXICAN AIDS PATIENT IN A BACULOVIRUS SYSTEM
Abstract
El objetivo principal del presente estudio fue la implementación de un sistema vector de expresión en baculovirus en células de insecto (IC-BECS), para la expresión de la proteína recombinante gp120 del virus de la inmunodeficiencia humana (VIH). Para esto, se aislaron secuencias de la proteína gp120 de una paciente mexicano con SIDA, y del plásmido de referencia HXB2, los cuales
se clonaron en forma independiente en el plásmido pFastBac1. Posteriormente, se utilizaron estos plásmidos para transformar E. coli DH10Bac conteniendo el bacmido bMON14272 del baculovirus AcMNPV, para luego inducir una transposición sitio dirigida para transferir el genoma de gp120 del VIH al bacmido del baculovirus. Se detectó el baculovirus recombinante de Bac-69-1 (paciente) y
el plásmido de referencia Bac-HX-4, utilizando la técnica de PCR y la fenotipificación de colonias. Después se encapsuló el genoma desnudo baculoviral en liposomas para su transfección al cultivo de células de insecto Sf9. Se utilizaron, además, análisis de Western blot (que se revelaron por quimioluminiscencia e hidrólisis de peróxido de hidrógeno) para la
identificación de la expresión de la proteína gp120. Los resultados obtenidos demostraron la generación de dos baculovirus recombinantes que expresaron la gp120 del paciente mexicano de SIDA o el plásmido de referencia. Se discute el potencial de utilizar a los baculovirus recombinantes de gp120 como vacunas del VIH en México.
Palabras claves: VIH, gp120, SIDA, pacientes Mexicanos, baculovirus, cultivo de células de insecto.
HIV, gp120, AIDS, Mexican patient, baculovirus, insect cell culture.
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