| Description |
Title: "Improving Recombineering beyond Escherichia coli MG1655 through the Broad Host-range Expression and Serial Evolutionary Enrichment of Metagenomic Library of Recombinases"
Committee:
George Church, Harvard (Research Advisor)
Jim Collins, BU BME (Academic Advisor)
Mo Khalil, BU BME
Wilson Wong, BU BME
Abstract:
This thesis project proposes to create a metagenomic library of recombinases, to test, characterize and demonstrate the functionality of its members in E.coli, and to expand it on the selected Gamma Proteobacteria close related to E.coli MG1655, including E.coli BL21, Salmonella enterica, Shigella flexneri, Vibrio cholera and Acinetobacter baylyi. We will also design and construct portable modules encoding the genetic elements required for successful expression and selection of the recombinase library. This genetic unit consisting of precursors and regulation module will provide with regulatory elements required for activation of silent gene clusters, and potentially, expression of precursor elements necessary for natural products. This module will be portable to multiple species, and it will be easily editable. By applying a broad host-range expression platform, we will be able to express the library in each individual organism. Recombination event, especially ssDNA recombination efficiency of individual recombinase will be studied. Competent candidate will be tested to improve recombineering efficiency in respective organism chassis. |
