Prospecting metagenomic enzyme subfamily genes for DNA family shuffling by a novel PCR-based approach.

ABSTRACT: DNA family shuffling is a powerful method for enzyme engineering, which utilizes recombination of naturally occurring functional diversity to accelerate laboratory-directed evolution. However, the use of this technique has been hindered by the scarcity of family genes with the required level of sequence identity in the genome database. We describe here a strategy for collecting metagenomic homologous genes for DNA shuffling from environmental samples by truncated metagenomic gene-specific PCR (TMGS-PCR). Using identified metagenomic gene-specific primers, twenty-three 921-bp truncated lipase gene fragments, which shared 64-99% identity with each other and formed a distinct subfamily of lipases, were retrieved from 60 metagenomic samples. These lipase genes were shuffled, and selected activ

SUBMITTER: Wang Q 

PROVIDER: S-EPMC3009877 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature