940 resultados para Vector insects


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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.

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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.

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Female choice is an important element of sexual selection that may vary among females of the same species. Few researchers have investigated the causes of variation in selectivity with respect to potential mates and overall level of motivation toward a stimulus source representative of a mate. This study demonstrates that female age may be one cause of variation in female choice. Females of different ages may have different mate preferences. As females age, they have less time left to reproduce, and their residual reproductive value decreases. This should correspond to a higher reproductive effort which may be represented as increased motivation and/or decreased selectivity. The effect of age on mate choice in Gryllus integer was investigated by using a non-compensating treadmill, called the Kugel, to measure female phonotaxis. Artificially generated male calling songs of varying pulse rates were broadcast in either a singlestimulus or a three-stimulus experimental design. The pulse rates used in the calling song stimuli were 70, 64, 76, 55 and 85 pulses per second. These corresponded to the documented mean pulse rate for the species at the experimental temperature, one standard deviation below and above the mean, and 2.5 standard deviations below and above the mean, respectively. Test females were either 11-14 days or 25-28 days post-ecdysis. Trials usually were conducted two to seven hours into the scotophase. In the single-stimulus experiment, females were presented with stimuli with only one pulse rate. Older females achieved higher vector scores than younger females, indicating that older females are more motivated to mate. Both groups showed little phonotactic response towards 55 or 85 pIs, both of which lie outside the natural range of G. integer calling song at the experimental temperature. Neither group discriminated among the three pulse rates that fell within the natural range of calling song. In the three-stimulus experiment, females were presented with stimuli with one of three pulse rates, 64, 70 or 76 pIs, In alternation. Both age groups had reduced responsiveness in this experiment, perhaps due to an increase in perceived male density. Additionally, younger females responded significantly more to 64 and 70 pIs than to the higher pulse rate, indicating that they are selective with respect to mate choice. Older females did not discriminate among the three pulse rates. Therefore, it was concluded that selectivity decreases with age. A further study was conducted to determine that these effects were due to age and not due to the differing periods without a mating between the two age groups. Again, stimuli were presented in a three-stimulus experimental design. Age was held constant at 28 days and time since last mating varied from 11 to 25 days. Females varyIng in time since last mating did not differ in their responses to the calling song pulse rates. This indicated that the increased motivation and decreased selectivity exhibited In the initial experiments were due to age and not to time without a mating. Neither time of trial nor female weight had an effect upon female phonotaxis. Data are discussed in terms of mate choice, residual reproductive value, and costs of choice.

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

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

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

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

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

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

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

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