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✔ Deep Sequencing of Systematic Combinatorial Libraries Reveals β-Lactamase Sequence Constraints at High Resolution

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Journal of Molecular Biology, Vol. 424, No. 3-4. (December 2012), pp. 150-167, doi:10.1016/j.jmb.2012.09.014
by Zhifeng Deng, Wanzhi Huang, Erol Bakkalbasi, et al.Nicholas G. Brown, Carolyn J. Adamski, Kacie Rice, Donna Muzny, Richard A. Gibbs, Timothy Palzkill
posted to no-tag by lynx ✚ on 2012-11-10 10:09:48 ** along with 2 people and 1 group barry GrayLab GrantLab
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Abstract

In this study, combinatorial libraries were used in conjunction with ultrahigh-throughput sequencing to comprehensively determine the impact of each of the 19 possible amino acid substitutions at each residue position in the TEM-1 β-lactamase enzyme. The libraries were introduced into Escherichiacoli, and mutants were selected for ampicillin resistance. The selected colonies were pooled and subjected to ultrahigh-throughput sequencing to reveal the sequence preferences at each position. The depth of sequencing provided a clear, statistically significant picture of what amino acids are ...
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