E. coli DH5α was used for construction of plasmids, and E. coli BL21 star(DE3)ΔlacZ was employed as the initial strain for metabolic engineering. The sequences of lgtA (GenBank accession number: AAK85138.1) from Neisseria meningitidis, Hpa-GalT (GenBank accession number: WP_049365321.1) from H. parainfluenzae and Aa-GalT (GenBank accession number: BAS48030.1) from A. actinomycetemcomitans NUM4039 were codon-optimized and synthesized by Tsingke Biotech Co., Ltd. (Beijing, China). All other genes were obtained from respective genomic DNA of E. coli MG1655 or BL21 star(DE3) using super-fidelity DNA polymerase.
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