| Title |
Tetramethylpyrazine-inducible promoter region from Rhodococcus jostii TMP1 / |
| Authors |
Stanislauskienė, Rūta ; Kutanovas, Simonas ; Kalinienė, Laura ; Bratčikov, Maksim ; Meškys, Rolandas |
| DOI |
10.3390/molecules23071530 |
| Full Text |
|
| Is Part of |
Molekules.. Basel : MDPI. 2018, vol. 23, iss. 7, art. no. 1530, p. [1-15].. ISSN 1420-3049 |
| Keywords [eng] |
Rhodococcus sp ; inducible promoter ; 2,3,5,6-tetramethylpyrazine ; LAL subfamily ; LuxR |
| Abstract [eng] |
An inducible promoter region, P-TTMP (tetramethylpyrazine [TTMP]), has been identified upstream of the tpdABC operon, which contains the genes required for the initial degradation of 2,3,5,6-tetramethylpyrazine in Rhodococcus jostii TMP1 bacteria. In this work, the promoter region was fused with the gene for the enhanced green fluorescent protein (EGFP) to investigate the activity of P-TTMP by measuring the fluorescence of bacteria. The highest promoter activity was observed when bacteria were grown in a nutrient broth (NB) medium supplemented with 5 mM 2,3,5,6-tetramethylpyrazine for 48 h. Using a primer extension reaction, two transcriptional start sites for tpdA were identified, and the putative -35 and -10 promoter motifs were determined. The minimal promoter along with two 15 bp long direct repeats and two 7 bp inverted sequences were identified. Also, the influence of the promoter elements on the activity of P-TTMP were determined using site-directed mutagenesis. Furthermore, P-TTMP was shown to be induced by pyrazine derivatives containing methyl groups in the 2- and 5-positions of the heterocyclic ring, in the presence of the LuxR family transcriptional activator TpdR. |
| Published |
Basel : MDPI |
| Type |
Journal article |
| Language |
English |
| Publication date |
2018 |
