Construction of an inducible expression shuttle vector for Laribacter hongkongensis, a novel bacterium associated
Patrick C Y Woo1, Shirley S L Ma, Jade L L Teng
1Department of Microbiology, Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital Compound, Pokfulam Road, Hong Kong.
Abstract:
An Escherichia coli-Laribacter hongkongensis shuttle vector (pPW380) was constructed by ligating the 4701-bp EcoRI digested fragment of pHLHK8 to EcoRI digested pBK-CMV. An E. coli-L. hongkongensis inducible expression shuttle vector was further constructed by ligating a 2105-bp fragment that contains the tetracycline repressor and tetracycline-inducible promoter region of pALC2084 to the 8897-bp fragment of pPW380, deletion of the green fluorescent protein gene, and insertion of a multiple cloning site. This inducible expression system was able to express two commonly used reporter genes, the green fluorescent protein gene and the glutathione S-transferase gene, efficiently in E. coli and L. hongkongensis.


