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Updated: Jul 13, 2026
CRISPR-based Shuttle Cloning: A High-throughput Cloning Method
Published on: June 13, 2025
Plasmid profile and construction of a small shuttle vector in Laribacter hongkongensis
Patrick C Y Woo1, Shirley S L Ma, Jade L L Teng
1State Key Laboratory of Emerging Infectious Diseases, Department of Microbiology, The University of Hong Kong, Room 423, University Pathology Building, Queen Mary Hospital Compound, Pokfulam, Hong Kong.
Abstract:
Among 21 human strains of Laribacter hongkongensis, small plasmids were observed in four strains, and large ones in six strains. The smallest, 3264-bp plasmid, pHLHK19, has only one ORF that encodes a putative replication initiator protein and a predicted origin of replication (ori) with a DnaA box, three 18-bp direct repeats and five pairs of inverted repeats. An Escherichia coli-L. hongkongensis shuttle vector was constructed by ligating the HindIII-digested pHLHK19, containing the replication initiator protein and ori of pHLHK19, to HindIII-digested pBK-CMV. This shuttle vector can propagate in E. coli and L. hongkongensis with good transformation efficiencies.
