Caracterização de biblioteca genômica enriquecida com regiões de microssatélites de Duroia macrophylla Huber (Rubiaceae)

Abstract

Apuruí, whose scientific name is Duroia macrophylla, belongs to the family Rubiaceae that stands out for the production of bioactive substances. In this sense, studies have verified biological activities presented by substances isolated from the specie. Action against Artemia salina, antitumor activity and activity against the causative agent of tuberculosis was verified. Although the biological activities presented, a species does not present genetic studies, so it becomes important to construct a genomic library enriched with regions of microsatellites, which are regions along the DNA consisting of small DNA sequences with 1 to 6 nucleotides in length, polymorphic and abundant tandem repeats. This marker can be classified into perfect, imperfect and composite sequences, and the number of nucleotides found in each replicate can be classified as: mono, di, tri, tetra, penta and hexanucleotides. Thus, the general objective of this work was to characterize a genomic library enriched with microsatellite regions for D. macrophylla. Initially, extraction of the genomic DNA was performed and, from this, the construction of the genomic library with 192 clones, from which, the plasmidial DNA extraction was performed. The nucleotide sequencing of the recombinant clones was performed on ABI 3130 xl automatic analyzer, and from the obtained nucleotide sequences it was possible to characterize the genomic library. After obtaining the microsatellites, as well as their characterization, the design of flanking primers was carried out to these regions. From the sequencing of the plasmid DNA of 192 recombinant clones, a total of 104 clones were obtained with good sequence quality, of these 13 had microsatellites to date. These were characterized as dinucleotides (30.77%), trinucleotides (38.46%), tetranucleotides (15.39%), pentanucleotides (7.69%) and hexanucleotides (7.69%) and as to the nature of repetition, all microsatellites (100%) were characterized as perfect. Microsatellite molecular markers have been developed in several studies, aiming to reach different responses, through the wide applicability of this marker, these studies were directed to different plant species, such as: palmetto, passion fruit, banana, among others.