RESUMO
This chapter describes the various methods derived from the protocol of standard fluorescent in situ hybridization (FISH) that are used in human, animal, plant, and microbial studies. These powerful techniques allow us to detect and physically map on interphase nuclei, chromatin fibers, or metaphase chromosomes probes derived from single-copy genes to repetitive DNA sequences. Other variants of the technique enable the co-localization of genes and the overall comparison of the genome among individuals of the same species or of different taxa. A further variant detects and localizes bacteria on tissues and cells. Overall, this offers a remarkable multiplicity of possible applications ranging from strict physical mapping, to clinical and evolutionary studies, making it a powerful and informative complement to other molecular, functional, or genomic approaches.
Assuntos
Análise Citogenética/métodos , Hibridização in Situ Fluorescente/métodos , Animais , Bactérias/metabolismo , Cromossomos Humanos/metabolismo , Cromossomos de Plantas/metabolismo , Hibridização Genômica Comparativa , DNA/metabolismo , Humanos , Sondas de Oligonucleotídeos/metabolismo , Plantas/metabolismo , Cariotipagem EspectralRESUMO
We have isolated and characterized AtREM1, the Arabidopsis ortholog of the cauliflower (Brassica oleracea) BoREM1. AtREM1 belongs to a large gene family of more than 20 members in Arabidopsis. The deduced AtREM1 protein contains several repeats of a B3-related domain, and it could represent a new class of regulatory proteins only found in plants. Expression of AtREM1 is developmentally regulated, being first localized in a few central cells of vegetative apical meristems, and later expanding to the whole inflorescence meristem, as well as primordia and organs of third and fourth floral whorls. This specific expression pattern suggests a role in the organization of reproductive meristems, as well as during flower organ development.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Meristema/genética , Sequência de Aminoácidos , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Sequência de Bases , DNA Complementar/química , DNA Complementar/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Meristema/crescimento & desenvolvimento , Dados de Sequência Molecular , Família Multigênica , Mutação , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Transcrição GênicaRESUMO
Meiotic associations at metaphase I have been analysed in Aegilops ovata × Triticum aestivum hybrid plants (genome constitution ABDUM) with low and high homoeologous pairing by using C-banding. Five different types of meiotic associations involving Aegilops and wheat genomes were identified. Pairing affinities between Aegilops and wheat genomes have been analysed from meiotic associations at metaphase I in low and high homoeologous pairing hybrid plants as well as from different meiotic configurations (bivalents and multivalents) in those hybrids with a high pairing mutant (phlb). Those kinds of distinguishable associations revealed the same relative order: AD-UM > A-D > U-M > AD-B > UM-B in both low and high homoeologous pairing hybrids. The mean number of associations per total associations (relative contribution) for the different distinguishable types of pairing was well maintained among hybrids with different levels of pairing (low and high) as well as in different meiotic configurations (bivalents and multivalents) in the high pairing plants. These results seem to indicate that the affinities expressed between the genomes that are in competition for pairing are independent of the meiotic configurations considered and on the level of pairing analysed.
