METHOD FOR TRACING GRAM-NEGATIVE BACTERIA INSIDE ANIMAL MODEL USING STABLE AND BIOLUMINESCENCE-BASED EXPRESSION SYSTEM THEREFOR

摘要

A method of creating a biotechnological product and an efficient and stable bio-luminescence vector which could be used for tracking Gram-negative bacteria when distributing inside animal body are provided. Through conjugation, this auto-luminescence vector can be easily transmitted from bacteria to bacteria among Gram-negative bacteria, and may facilitate bacteria to be luminescence-labeled for subsequently analyzing the dynamic change of bio-luminescent bacteria within animal body in vivo. This system includes a lacZ promoter-driven IuxABCDE, a high copy number of ColE1 replicon, and a high plasmid stability of the conjugative and broad host-ranged plasmid pSE34 from Salmonella enterica serovar Enteritidis Sal550. This resulting construct pSE-Lux1 can not only conjugatively transmit among bacteria with broad host range, but also stably maintain in bacteria to efficiently express the bio-luminescent IuxABCDE without supplementing the subtract for luciferases and the antibiotics for plasmid selection.