Construction of an inducible expression shuttle vector for Laribacter hongkongensis, a novel bacterium associated with gastroenteritis.
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.
Woo
Patrick C Y
PC
Department of Microbiology, Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital Compound, Pokfulam Road, Hong Kong.
Ma
Shirley S L
SS
Teng
Jade L L
JL
Li
Maria W S
MW
Kao
Richard Y T
RY
Lau
Susanna K P
SK
Yuen
Kwok-Yung
KY
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005759:Gastroenteritis / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009346:Neisseriaceae / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016870:Neisseriaceae Infections / Q000382:microbiology; D010957:Plasmids; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase