| description |
Metagenomics allow us to screen the genic content from, virtually, any type of environment with biological content and rendering the possibility of discovering products of interest. Here we propose a method to obtain medium-insert-size (7-15 Kb) metagenomic libraries that are sequenced through a hybrid approach based on pyrosequencing of clone pools and later Sanger-end sequencing of individual clones. Sequences are then assembled and the open reading frames annotated. We have applied this methodology to study human gut microbiota. From the 358 analysed clones from the library obtained from a faecal sample we found various ORFs annotated as enzymes with already reported industrial or medical application. Finally, forty-three ORFs were annotated as proteins of still unknown function. This method represents a new approach to explore microbial ecosystems, particularly the human gut microbiome, providing a scalable access to clones with potentially biotechnological and biomedical applications. Thus, previous sequence knowledge will drive the choice of the correct functional screening approach. |