Please use this identifier to cite or link to this item: https://dspace.ctu.edu.vn/jspui/handle/123456789/71747
Title: Development of inducer-free expression plasmids using IPTG-inducible Pspac promoter for Bacillus subtilis
Authors: Chu, Thi Bich Phuong
Phan, Hanh Thi Thu
Nguyen, Hoang Duc
Phan, Trang Thi Phuong
Keywords: Bacillus subtilis
Inducer-free
pHT vector
Pspac
Weak promoter
Issue Date: 2021
Series/Report no.: Vietnam Journal of Science, Technology and Engineering;Vol. 63, No. 01 .- P.64-70
Abstract: An inducer-free expression vector for the low expression levels in Bacillus subtilis (B. subtilis) is necessary for fundamental research. In this study, we constructed inducer-free expression plasmids carrying Pspac, a well-known IPTG-inducible promoter, by removing a part of the lad gene. Then, we analysed the expression of the target genes bgaB and gfp* in B. subtilis. Western blot experiments demonstrated that the reporters from the inducer-free plasmids with Pspac could be produced at low levels in B. subtilis strains and were equivalent to their corresponding inducible constructs with 1 mM IPTG. The reporter activities showed that inducer-free expression from the Pspac promoter was dramatically less than that of the inducer-free plasmids with the strong promoters Pgrac01 and Pg/m100 reported previously, about 16.2 to 20.3 times for BgaB and 24.7 to 34.3 times for GFP⁺, respectively. In conclusion, the inducer-free expression vectors carrying Pspac promoters allow the constitutive expression of heterologous recombinant proteins at low levels in B. subtilis.
URI: https://dspace.ctu.edu.vn/jspui/handle/123456789/71747
ISSN: 2525-2461
Appears in Collections:Vietnam journal of Science, Technology and Engineering

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