Table 1.
List of sequences of oligonucleotide primers, plasmids and strains used
| Name | Sequence (5′–3′) or function | Relevant characteristics or reference |
|---|---|---|
| A. Oligonucleotides | ||
| xylB_fw_Bsu | GAGAAAGGAGGCCCTTCAGATGAAGTATGTCATTGGAATAGATCTTGG | HE of Bsu xylB; RBS; start |
| xylB_rv_Bsu | GATCTAGATTAGTTTTTTCGAAAGCTCTTCAAAGC | HE of Bsu xylB; XbaI; stop |
| xylB_fw_Cgl | GAGAAAGGAGGCCCTTCAGATGGCTTTGGTTCTTGGAATCG | OE of Cgl xylB; RBS; start |
| xylB_rv_Cgl | GATCTAGACTAGTACCAACCCTGCGTTG | OE of Cgl xylB; XbaI; stop |
| xylB_fw_Eco | GAGAAAGGAGGCCCTTCAGATGTATATCGGGATAGATCTTGGCAC | HE of Eco xylB; RBS; start |
| xylB_rv_Eco | GATCTAGATTACGCCATTAATGGCAGAAGTTG | HE of Eco xylB; XbaI; stop |
| xylA_fw_Bsu | GATCTAGAGAAAGGAGGCCCTTCAGATGGCTCAATCTCATTCCAGTTCA | HE of Bsu xylA; XbaI; RBS; start |
| xylA_rv_Bsu | GAGAGCTCTTATACTTCTAAAATGTATTGGTTCAATATCGCT | HE of Bsu xylA; Ecl136II; stop |
| xylA_fw_Eco | GATCTAGAGAAAGGAGGCCCTTCAGATGCAAGCCTATTTTGACCAGC | HE of Eco xylA; XbaI; RBS; start |
| xylA_rv_Eco | GAGAGCTCTTATTTGTCGAACAGATAATGGTTTACCAG | HE of Eco xylA; Ecl136II; stop |
| xylA_fw_Msm | GATCTAGAGAAAGGAGGCCCTTCAGATGACCGTGTTGGAGTCGAA | HE of Msm xylA; XbaI; RBS; start |
| xylA_rv_Msm | GAGAGCTCTCATCGCGCGCCCATCAG | HE of Msm xylA; Ecl136II; stop |
| xylA_fw_Xcc | GATCTAGAGAAAGGAGGCCCTTCAGATGAGCAACACCGTTTTCATCG | HE of Xcc xylA; XbaI; RBS; start |
| xylA_rv_Xcc | GAGAGCTCTCAACGCGTCAGGTACTGATT | HE of Xcc xylA; Ecl136II; stop |
| B. Plasmids | ||
| pEKEx3 | SpecR; C. glutamicum/E. coli shuttle vector (Ptac, lacIq; pBL1, OriVCg, OriVEc) | Stansen et al. (2005) |
| pEKEx3-xylAEc | Derived from pEKEx3, for regulated expression of xylAEc (b3565) of E. coli | Gopinath et al. (2011) |
| pEKEx3-xylABs | Derived from pEKEx3, for regulated expression of xylABs (BSU17600) of B. subtilis | This work |
| pEKEx3-xylAMs | Derived from pEKEx3, for regulated expression of xylAMs (MSMEG_6021) of M. smegmatis | This work |
| pEKEx3-xylAXc | Derived from pEKEx3, for regulated expression of xylAXc (XCC1758) of X. campestris | This work |
| pEKEx3-xylAXc-xylBEc | Derived from pEKEx3, for regulated expression of xylAXc (XCC1758) of X. campestris and xylBEc (b3580) of E. coli | This work |
| pEKEx3-xylAXc-xylBBs | Derived from pEKEx3, for regulated expression of xylAXc (XCC1758) of X. campestris and xylBBs (BSU17610) of B. subtilis | This work |
| pEKEx3-xylAXc-xylBCg | Derived from pEKEx3, for regulated expression of xylAXc (XCC1758) of X. campestris and xylBCg (cg0147) of C. glutamicum | This work |
| pVWEx1 | KanR; C. glutamicum/E. coli shuttle vector (Ptac, lacIq; pHM1519, OriVCg, OriVEc) | Peters-Wendisch et al. (1998) |
| pVWEx1-araBAD | Derived from pVWEx1, for regulated expression of araB (b0063) and araA (b0062) and araD (b0061) of E. coli | Schneider et al. (2011) |
| C. Strains | ||
| E. coli | ||
| DH5α | Fthi-1 endA1 hsdr17(r−, m−) supE44 _lacU169 (ф80lacZ_M15) recA1 gyrA96 relA1 | Hanahan (1983) |
| C. glutamicum | ||
| ATCC13032 | Wild type (WT) | Kinoshita et al. (1957) |
| DM1729 | lysC P458S, homV59A, pyc T311I | Georgi et al. (2005) |
| ORN1 | l-ornithine overproducing strain derived from ATCC13032, auxotrophic for l-arginine due to argF deletion | Schneider et al. (2011) |
| PUT21 | ORN1 carrying pVWEx1-speC-5′21-argF | Schneider et al. (2011) |
Restriction sites are underlined, ribosomal binding sites are shown in bold, stop and start codons are in italics.
OE, overexpression; HE, heterologous expression; RBS, ribosomal binding site; Cgl, C. glutamicum; Eco, E. coli; Bsu, B. subtilis; Msm, M. smegmatis; Xcc, X. campestris.