RESUMO
C. elegans is a well-characterized and relatively simple model organism, making it attractive for studying nuclear pore complex proteins in cell and developmental biology. C. elegans is transparent and highly amendable to genetic manipulation. Therefore, it is possible to generate fluorescently tagged proteins and combine this with various light microscopy techniques to study protein behavior in space and time. Here, we provide protocols to prepare both fixed and live C. elegans for confocal and light sheet microscopy. This enables the analysis of nuclear pore complex proteins from embryonic stages to the aging adult.
Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Complexo de Proteínas Formadoras de Poros Nucleares , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Microscopia de Fluorescência/métodos , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismoRESUMO
Nineteen microsatellite loci developed for the Eurasian otter (Lutra lutra) and 15 loci developed for the North American river otter (Lontra canadensis) were tested for ease of amplification and degree of polymorphism on a set of 20 giant otter (Pteronura brasiliensis) faecal samples from the Bolivian Amazon basin. Nineteen loci amplified consistently well, with polymorphisms ranging from two to nine alleles and observed heterozygosity ranging from 0.15 to 0.85.