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Published on: June 25, 2015
Strong synthetic promoters in Mycobacteria
Junjie Wen1, Yongmei Zhang2, Luwei Cao3
1Shanghai Key Laboratory of Infectious Diseases and Biosafety Emergency Response, National Medical Center for Infectious Diseases, Huashan Hospital, Shanghai, China; School of life science, Fudan University, Shanghai, China.
None:
Finding potent promoters that would enable foreign genes to be expressed at high levels in Mycobacteria would be extremely helpful in the creation of recombinant vaccines and features for Mycobacterial antigens. Recombinant mycobacterial gene expression is commonly carried out using Mycobacterium smegmatis (M. smegmatis) as the host. In this paper, we constructed a promoter library with minimized consensus promoter sequence of mycobacteria. This library was ligated to the upstream of green fluorescent protein (EGFP) and transformed into M. smegmatis. Strength of the promoters was revealed by the green fluorescent of the colonies under a blue LED transilluminator. The strong fluorescent colonies were picked up and the promoter sequence was analyzed by sanger sequencing. Overall, we got plenty of strong promoters less than 100 base pair, which can be cloned into other plasmids easily, and would be a useful tool in Mycobacteria genetics.
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