Characterization of Protein Domain Function via in vitro DNA Shuffling

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

Detail(s)

Original languageEnglish
Article numberARTN e2873
Journal / PublicationBio-protocol
Volume8
Issue number11
Publication statusPublished - 5 Jun 2018
Externally publishedYes

Abstract

We recently investigated the molecular events that drive evolution of the CTX-M-type beta-lactamases by DNA shuffling of fragments of the bla(CTX-M-14) and bla(CTX-M-15) genes. Analysis of a total of 51 hybrid enzymes showed that enzymatic activity could be maintained in most cases, yet the enzymatically active hybrids were found to possess much fewer amino acid substitutions than the few hybrids that became inactive, suggesting that point mutations in the constructs rather than reshuffling of the fragments of the two target genes would more likely cause disruption of CTX-M activity. Certain important residues that played important functional roles in mediating enzyme activity were identified. These findings suggest that DNA shuffling is an effective approach to identify and characterize important functional domains in bacterial proteins.

Research Area(s)

  • CTX-M-14, CTX-M-15, DNA shuffling, Hybrid enzyme, Evolution, HYBRID, GENES

Citation Format(s)

Characterization of Protein Domain Function via in vitro DNA Shuffling. / Po, Kathy Hiu Laam; Chan, Edward Wai Chi; Chen, Sheng.

In: Bio-protocol, Vol. 8, No. 11, ARTN e2873, 05.06.2018.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review