Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Tropaire-prosier à l'usage de Moissac, avec notation en neumes.

Provient de la bibliothèque du chanoine Morelot, de Dijon, et a été publié sous le titre de Tropaire-prosier de l'abbaye de Saint-Martin de Montauriol, par l'abbé C. Daux (Paris, 1901, in-8°) ; cf. l'art. du Rév. H. M. Bannister, dans la Revue d'histoire et de littérature religieuses (1903), t. VIII, p. 556 et suiv.

Development of a bovine adenovirus type 2-based gene delivery vector /

ABSTRACT Recombinant adenoviruses are currently under intense investigation as potential gene delivery and gene expression vectors with applications in human and veterinary medicine. As part of our efforts to develop a bovine adenovirus type 2 (BAV2) based vector system, the nucleotide sequence of BAV2 was determined. Sixty-six open reading frames (ORFs) were found with the potential to encode polypeptides that were at least 50 amino acid (aa) residue long. Thirty-one of the BAV2 polypeptide sequences were found to share homology to already identified adenovirus proteins. The arrangement of the genes revealed that the BAV2 genomic organization closely resembles that of well-characterized human adenoviruses. In the course of this study, continuous propagation of BAV2 over many generations in cell culture resulted in the isolation of a BAV2 spontaneous mutant in which the E3 region was deleted. Restriction enzyme, sequencing and PCR analyses produced concordant results that precisely located the deletion and revealed that its size was exactly 1299 bp. The E3-deleted virus was plaque-purified and further propagated in cell culture. It appeared that the replication of such a virus lacking a portion of the E3 region was not affected, at least in cell culture. Attempts to rescue a recombinant BAV2 virus with the bacterial kanamycin resistance gene in the E3 region yielded a candidate as verified with extensive Southern blotting and PCR analyses. Attempts to purify the recombinant virus were not successful, suggesting that such recombinant BAV2 was helper-dependent. Ten clones containing full-length BAV2 genomes in a pWE15 cosmid vector were constructed. The infectivity of these constructs was tested by using different transfection methods. The BAV2 genomic clones did appear to be infectious only after extended incubation period. This may be due to limitations of various transfection methods tested, or biological differences between virus- and E. co//-derived BAV2 DNA.

Respuesta funcional de Toxorhynchites theobaldi (DYAR y KNAB) sobre larvas de Aedes aegypti (LINN.) vector del dengue

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) UANL

Control del vector del dengue Aedes aegypti (l) con la participación de estudiantes de primaria como educadores de salud en la cuidad de Guadalupe Nuevo Léon

Tesis (Maestria en Ciencias con Especialidad en Entomología Médica) UANL

Comparación de la eficiencia de rociados domiciliares en ultra bajo volumen (ULV) y termonebulización para el control de poblaciones adultas del vector del dengue Aedes aegypti (L.) en una colonia de Guadalupe, Nuevo León, México

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) UANL

Bionomia de Aedes aegypti L., vector del dengue en el sur del estado de Jalisco, México

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) UANL

Bionomía del vector de la malaria Anopheles vestitipennis Dyar & Knab (Díptera: Culicidae) en la cuenca del Río Polochic, Alta Verapaz, Guatemala, C.A. [por] Jaime Abraham Juárez Sandoval

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) U.A.N.L.

Evaluación en campo del depredador Mesocyclops longisetus (Thiebaud) (Copepoda: Cyclopoidea) para el control larval del vector del dengue Aedes aegypti (L.) en Monterrey, Nuevo León [por] Norma Gorrochotegui Escalante

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) U.A.N.L.

Construcción del vector adenoviral AD-UV2

Tesis (Maestría en Ciencias con Especialidad en Biología Molecular e Ingeniería Genética) UANL, 2011.

Generación del vector adenoviral oncolítico Ad5

Tesis (Maestría en Ciencias con Especialidad en Biología Molecular e Ingeniería Genética) UANL, 2011.

Generación de un vector adenoviral para la producción de anticuerpos policlonales contra la proteína ENV del XMRV (Xenotropic murine leukemia virus-related virus).

Tesis (Maestría en Ciencias con Orientación en Biología Molecular e Ingeniería Genética) UANL, 2011.

A composite discrete traction vector.

Tesis (Maestría en Ciencias con Orientación en Ingeniería Estructural) UANL, 2013.

Generación de un vector de recombinación homóloga para la inactivación condicional del gen NURR 1 .html Humberto Rodríguez Rocha.

Tesis (Doctor en Ciencias con Orientación Terminal en Morfología) U.A.N.L., 2006.