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Efficient secretion of a modified E7 protein from human papilloma virus type‐16 by Lactococcus lactis
Authors:D Quistián‐Martínez  J Villatoro‐Hernández  MJ Loera‐Arias  BR Rangel‐Colmenero  LM Zavala‐Flores  J Sepúlveda‐Saavedra  S Guzmán‐López  RE Elizondo‐Omaña  R Montes‐de‐Oca‐Luna  O Saucedo‐Cárdenas
Institution:1. Departamento de Bioquímica, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León (UANL), San Nicolás de los Garza, N.L. México;2. Departamento de Histología, Facultad de Medicina, UANL, Monterrey, N.L. México;3. Departamento de Anatomía Humana, Facultad de Medicina, UANL, Monterrey, N.L. México;4. División de Genética, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social (IMSS), Monterrey, N.L. México
Abstract:Aims: To create and provide a strain of the food‐grade bacterium Lactococcus lactis able to efficiently secrete a modified form of the E7 protein from the human papilloma virus (HPV) type‐16. Methods and Results: We cloned the coding sequence of a modified E7 (E7m) from the HPV‐16 in a plasmid regulated by the strong expression promoter p59. Secretion of the E7m was made by the signal peptide of the usp45 gene. The E7m was detected by Western blot in the cell‐free‐medium fraction, showing no degradation or aberrant forms. Conclusions: We constructed a strain of L. lactis able to secrete efficiently a HPV‐16 E7 modified protein with diminished transforming activity. Significance and Impact of the Study: Human papilloma virus infection is associated with more than 99% of cervical cancers. Immunotherapy targeting E7 to treat HPV‐associated cervical malignancies has been demonstrated to be highly efficient. However, native E7 maintains transforming activity. We present this new strain of a food‐grade bacterium able to efficiently secrete a HPV‐16 E7‐modified protein with diminished transforming activity. This new strain could be used as a live vaccine to deliver E7 at a mucosal level and generate antitumour immune responses against HPV‐associated tumours.
Keywords:antitumour protection  HPV‐16  Lactococcus lactis  modified E7  secretion
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