Construction and maintenance of randomized retroviral expression libraries for transmembrane protein engineering.

ABSTRACT: Genetic selection from libraries expressing proteins with randomized amino acid segments is a powerful approach to identify proteins with novel biological activities. Here, we assessed the utility of deep DNA sequencing to characterize the composition, diversity, size and stability of such randomized libraries. We used 454 pyrosequencing to sequence a retroviral library expressing small proteins with randomized transmembrane domains. Despite the potential for unintended random mutagenesis during its construction, the overall hydrophobic composition and diversity of the proteins encoded by the sequenced library conformed well to its design. In addition, our sequencing results allowed us to calculate a more accurate estimate of the number of different proteins encoded by the library and sugg

SUBMITTER: Marlatt SA 

PROVIDER: S-EPMC3038463 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature