INTEGRATED SYSTEM FOR HIGH THROUGHPUT CAPTURE OF GENETIC DIVERSITY

摘要

Compositions and methods for rapid and highly efficient characterization of genetic diversity in organisms are provided. The methods involve rapid sequencing and characterization of extrachromosomal DNA, particularly plasmids, to identify and isolate useful nucleotide sequences. The method targets plasmid DNA and avoids repeated cloning and sequencing of the host chromosome, thus allowing one to focus on the genetic elements carrying maximum genetic diversity. The method involves generating a library of extrachromosomal DNA clones, sequencing a portion of the clones, comparing the sequences against a database of existing DNA sequences, using an algorithm to select said novel nucleotide sequence based on the presence or absence of said portion in a database, and identification of at least one novel nucleotide sequence. The DNA sequence can also be translated in all six frames and the resulting amino acid sequences can be compared against a database of protein sequences. The integrated approach provides a rapid and efficient method to identify and isolate useful genes. Organisms of particular interest include, but are not limited to bacteria, fungi, algae, and the like. Compositions comprise a mini-cosmid vector comprising a stuffer fragment and at least one cos site.