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CHLOROPLAST GENETIC TOOL FOR THE GREEN MICROALGAE HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE,VOLVOCALES)1
Authors:Carla L Gutiérrez  Carolina Escobar  Sergio H Marshall  Vitalia Henríquez
Institution:1. Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Facultad de Ciencias. Pontificia Universidad Católica de Valparaíso. Avenida Universidad 330, Campus Curauma, Valparaíso, Chile;2. Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Facultad de Ciencias. Pontificia Universidad Católica de Valparaíso. Avenida Universidad 330, Campus Curauma. Valparaíso, Chile CREAS, Centro Regional de Alimentos Saludables, Valparaíso, Chile NBC, Núcleo de Biotecnología Curauma, Curauma, Valparaíso, Chile
Abstract:At present, there is strong commercial demand for recombinant proteins, such as antigens, antibodies, biopharmaceuticals, and industrial enzymes, which cannot be fulfilled by existing procedures. Thus, an intensive search for alternative models that may provide efficiency, safety, and quality control is being undertaken by a number of laboratories around the world. The chloroplast of the eukaryotic microalgae Haematococcus pluvialis Flotow has arisen as a candidate for a novel expression platform for recombinant protein production. However, there are important drawbacks that need to be resolved before it can become such a system. The most significant of these are chloroplast genome characterizations, and the development of chloroplast transformation vectors based upon specific endogenous promoters and on homologous targeting regions. In this study, we report the identification and characterization of endogenous chloroplast sequences for use as genetic tools for the construction of H. pluvialis specific expression vectors to efficiently transform the chloroplast of this microalga via microprojectile bombardment. As a consequence, H. pluvialis shows promise as a platform for expressing recombinant proteins for biotechnological applications, for instance, the development of oral vaccines for aquaculture.
Keywords:aadA gene  chloroplast transformation  Expression vector  5′  and 3′  rbcL regulatory sequences
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