RESUMO
OBJECTIVE: To establish a fast and simple method for screening mutant yeast strains. MATERIALS AND METHODS: Homologous recombination technique was used to detect mutant yeast strains in yeast genomic library, with the green fluorescent protein gene as the reporter gene in the transposon. RESULTS: The strains that emitted green fluorescence were isolated, indicating that the gfp gene was inserted into the yeast genome by homologous recombination. CONCLUSION: This study established a useful method for functional genome study by homologous recombination technique, and provide an alternative for gene therapeutic drug development.
Assuntos
Genes Fúngicos/genética , Proteínas de Fluorescência Verde/genética , Mutação , Recombinação Genética , Leveduras/genética , Clonagem Molecular/métodos , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Fluorescência Verde/análiseRESUMO
OBJECTIVE: To evaluate the relationship between receptor-mediated endocytosis, autophagosome formation and apoptosis by observing the morphological changes of mouse peritoneal macrophages in the course of receptor-mediated endocytotic activities, autophagy and apoptosis. METHODS: Mouse peritoneal macrophages were incubated with horseradish peroxidase-labeled concanavalin A (ConA-HRP), and morphological examinations were performed at different time points after the incubation. RESULTS: After the incubation of the macrophages with ConA-HRP, ConA-HRP was observed to enter the vesicles by way of receptor-mediated endocytosis. Three kinds of endosomes were observed, namely vesicles, tubes and double-membrane sheets. The double-membrane sheets enveloped a portion of the cytoplasm and organelles, thus giving rise to vesicles, or the autophagosomes, which later fused with lysosome, followed by the apoptosis of the macrophages. CONCLUSION: Receptor-mediated endocytosis of ConA-HRP is associated with autophagy and apoptosis.
