Improved Transposase Compositions for Whole Genome Sequencing
Posted Nov 05 2012 7:00pm
Description of Invention:
The invention provides improved transposase enzymes engineered to exhibit reduced sequence biases, and to operate more efficiently than wildtype transposases.
Scientists at NIDDK and John Hopkins University jointly developed mutant transposases that are superior to wildtype transposases in whole genome sequencing applications. Transposases facilitate the cleavage of certain DNA segments, called transposons, at specific sites within a genome and their subsequent insertions at random sites. Addition of transposases and labeled transposons to whole genome preparations allow for one-pot, simultaneous fragmentation and identification of targeted DNA sequences.
Mutations introduced by the inventors facilitate formation of dimeric enzyme complexes with enhanced activity and stability. These modifications result in more efficient fragmentation and tagging of genomic DNA.
Applications: Kits for whole genome sequencing.
Can easily be expressed in the bacterium, E. coli, and purified in large quantities.
Are soluble, stable and exist as smaller active complexes compared to native enzymes.
Are fully active at room temperature (23 - 30°C).
Have a higher transposition activity and show minimal insertional sequence bias in-vitro compared to the wild type.
In vitro data available
Inventors: Nancy L Craig (Johns Hopkins School of Medicine) Sunil Gangadharan (Johns Hopkins School of Medicine) Frederick Dyda (NIDDK) Allison B Hickman (NIDDK)