Development of inducer-free expression plasmids using IPTG-inducible Pspac promoter for Bacillus subtilis
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 lacI gene. Then, w...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Vietnam Ministry of Science and Technology
2021-03-01
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| Series: | Vietnam Journal of Science, Technology and Engineering |
| Subjects: | |
| Online Access: | https://vietnamscience.vjst.vn/index.php/vjste/article/view/22 |
| Summary: | 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 lacI 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 Pgrac100 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.
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| ISSN: | 2525-2461 2615-9937 |