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Research Article Review Jmb J. Microbiol. Biotechnol. (2017), 27(0), 1–1 https://doi.org/10.4014/jmb.1805.05013 Research Article Review jmb Supplementary Table S1. Bacterial strains used in this study. Strain Relevant characteristics* Source C. glutamicum ATCC 13032 Wild-type strain [1] C. glutamicum CP L-Leucine-producing, Ile, 2-TAr, SGr, LeuHxr, NVr This study C. glutamicum XV L-Valine-producing, Met, Ile, PA, 2-TAr, SGr, NVr This study XV C. glutamicum AN01 C. glutamicum ATCC 13032 ΔCgl2610 ::Ptuf+ilvBN This study XV C. glutamicum AN02 C. glutamicum AN01 ΔCgl1890 ::Ptuf+ilvBN This study C. glutamicum AN03 C. glutamicum AN02 ΔCgl2103 ::Ptuf+CP_103 This study *: Ile: L-isoleucine; Leu: L-leucine; LeuHx: L-leucine hydroxamate; Met: L-methionine; NV: norvaline; PA: pantoate; SG: sulfaguanidine; 2-TA: 2-thiazolylalanine. 1. Kalinowski J, Bathe B, Bartels D, Bischoff N, Bott M, Burkovski A, et al. 2003. The complete Corynebacterium glutamicum ATCC 13032 genome sequence an impact on the production of L-aspartate-derived amino acids and vitamins. J. Biotechnol. 104: 5-25. A 2017 ⎪ Vol. 27⎪ No. 0 Supplementary Table S2. Plasmids used in this study. Plasmid Relevant characteristics* Source r pXMJ19 Cm ; Ptac promoter [1] r pEC-XK99E Km ; Ptrc promoter [2] pK18mobsacB Kmr; sacB [3] r pXT01 Cm ; pXMJ19 ΔPtac::Ptuf This study pXT01+ilvBN ATCC 13032 Cmr; ATCC 13032 derived ilvBN in pXT01 This study pXT01+ilvBN CP Cmr; CP derived ilvBN in pXT01 This study pXT01+ilvBN XV Cmr; XV derived ilvBN in pXT01 This study pXT01+leuBCD Cmr; ATCC 13032 derived leuBCD in pXT01 This study pEC-XK99E+leuA ATCC 13032 Kmr; ATCC 13032 derived leuA in pEC-XK99E This study pEC-XK99E+leuA CP Kmr; CP derived leuA in pEC-XK99E This study pEC-XK99E+leuA XV Kmr; XV derived leuA in pEC-XK99E This study pEC-XK99E+leuA_2 CP Kmr; CP derived leuA_2 in pEC-XK99E This study r XV pK18mobsacB-Cgl2610 Km ; integrated Ptuf+ilvBN to Cgl2610 of ATCC 13032 This study r XV pK18mobsacB-Cgl1890 Km ; integrated Ptuf+ilvBN to Cgl1890 of ATCC 13032 This study r pK18mobsacB-CP_103 Km ; integrated Ptuf+CP_103 to Cgl2103 of ATCC 13032 This study pXT01+CP_49 Cmr; CP derived CP_49 in pXT01 This study pXT01+CP_103 Cmr; CP derived CP_103 in pXT01 This study pXT01+CP_570 Cmr; CP derived CP_570 in pXT01 This study pXT01+CP_760 Cmr; CP derived CP_760 in pXT01 This study pXT01+CP_788 Cmr; CP derived CP_788 in pXT01 This study pXT01+CP_967 Cmr; CP derived CP_967 in pXT01 This study pXT01+CP_1509 Cmr; CP derived CP_1509 in pXT01 This study pXT01+CP_1823 Cmr; CP derived CP_1823 in pXT01 This study pXT01+CP_1834 Cmr; CP derived CP_1834 in pXT01 This study pXT01+CP_2613 Cmr; CP derived CP_2613 in pXT01 This study pXT01+CP_2675 Cmr; CP derived CP_2675 in pXT01 This study pXT01+CP_3087 Cmr; CP derived CP_3087 in pXT01 This study pXT01+CP_3110 Cmr; CP derived CP_3110 in pXT01 This study pXT01+XV_130 Cmr; XV derived XV_130 in pXT01 This study pXT01+XV_1076 Cmr; XV derived XV_1076 in pXT01 This study pXT01+XV_1077 Cmr; XV derived XV_1077 in pXT01 This study pXT01+XV_1088 Cmr; XV derived XV_1088 in pXT01 This study pXT01+XV_1267 Cmr; XV derived XV_1267 in pXT01 This study pXT01+XV_2014 Cmr; XV derived XV_2014 in pXT01 This study pXT01+XV_2617 Cmr; XV derived XV_2617 in pXT01 This study pXT01+XV_2620 Cmr; XV derived XV_2620 in pXT01 This study pXT01+XV_2682 Cmr; XV derived XV_2682 in pXT01 This study pXT01+XV_2797 Cmr; XV derived XV_2797 in pXT01 This study *: Cm: chloramphenicol; Km: kanamycin. 1. Jakoby M, Ngouoto-Nkili CE, Burkovski A. 1999. Construction and application of new Corynebacterium glutamicum vectors. Biotechnol. Tech. 13: 437-441. 2. Kirchner O, Tauch A. 2003. Tools for genetic engineering in the amino acid-producing bacterium Corynebacterium glutamicum . J. Biotechnol. 104: 287-299. 3. Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G, Pühler A. 1994. Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum . Gene. 145: 69-73. Supplementary Table S3. Primers used in this study. Primer no. Target gene Sequence (5'-3') 1 Cgl2610 CTATGACATGATTACGAATTCAAGCGTGGCAACAACTGGAA 2 TTTCCTTCGGATCTAAACGATCTTCACCATCTCGCAGTAACCAGAG 3 ATGGCTCCGGCCAAGATCTAACTGAAGGCTGTGGTGTTTAACAAC 4 ACGACGGCCAGTGCCAAGCTTGGAAATGTTGGCAGTTAAGCG 5 Ptuf + ilvBN CTCTGGTTACTGCGAGATGGTGAAGATCGTTTAGATCCGAAGGAAAAC 6 GTTGTTAAACACCACAGCCTTCAGTTAGATCTTGGCCGGAGCCAT 7 Cgl1890 CTATGACATGATTACGAATTCATGCGTTCAGATGTTATCGAGTTACC 8 CCTTCGGATCTAAACGATCTCGCCAGTCAATGGGTTGTAGTAGT 9 ATGGCTCCGGCCAAGATCTAATACAGGAATGAAGGCTGGGGAT 10 ACGACGGCCAGTGCCAAGCTTTCATATGCGAAGCTTATTATAAAGATTC 11 Ptuf + ilvBN ACTACTACAACCCATTGACTGGCGAGATCGTTTAGATCCGAAGGAAAAC 12 ATCCCCAGCCTTCATTCCTGTATTAGATCTTGGCCGGAGCCAT 13 ilvBN CGAAGTCCAGGAGGAAAGCTTGTGAATGTGGCAGCTTCTCAACA 14 GCTGAATTCGAGCTCGGTACCTTAGATCTTGGCCGGAGCC 15 leuA AGGAAACAGACCATGGAATTCCGCACTACTGGTGATGATTGTTTC 16 CTTGCATGCCTGCAGGTCGACGGGAGGCGTCGTAAAGCTTAA 17 leuB CGAAGTCCAGGAGGAAAGCTTTGAGAAGGGAATCCCACAACA 18 CCTCAATGTCATCGCAGAACAGCAATCCAGCAAGTATTTACACCAAC 19 leuCD GTTGGTGTAAATACTTGCTGGATTGCTGTTCTGCGATGACATTGAGG 20 CGGGGATCCTCTAGAGTCGACTTAAGCGTTAGTGCGTGGCTT 21 CP_49 CGAAGTCCAGGAGGAAAGCTTGTGCCTTTTGACGTTTCATTCA 22 GCTGAATTCGAGCTCGGTACCTCACGCGGTGAAGTTTAACCT 23 CP_103 CGAAGTCCAGGAGGAAAGCTTATGAGCAGCTCAACACTTCTCCT 24 GCTGAATTCGAGCTCGGTACCTTATGCGCTGAGTGGCACG 25 CP_570 CGAAGTCCAGGAGGAAAGCTTATGCAGTTATCTCCTTCAAATACGTC 26 GCTGAATTCGAGCTCGGTACCCTAAAACTCCTTCTGGCTGCGC 27 CP_760 CGAAGTCCAGGAGGAAAGCTTATGAATAAACAGTCCGCTGCAGT 28 GCTGAATTCGAGCTCGGTACCCTAGGTGTGTGTACTCGCCTCTTC 29 CP_788 CGAAGTCCAGGAGGAAAGCTTATGAACGAGATGATTCTCGCCG 30 GCTGAATTCGAGCTCGGTACCTTAGCGGCGCATGATCTTCT 31 CP_967 CGAAGTCCAGGAGGAAAGCTTGTGGATAAAGATATTGAAAACCGCA 32 GCTGAATTCGAGCTCGGTACCTCACAGTCTGCCCTTTCTGAGG 33 CP_1509 CGAAGTCCAGGAGGAAAGCTTATGTCGCAACTGATGATTGATGC 34 GCTGAATTCGAGCTCGGTACCTTAGAGCAATGAAGACAGGAACTCTT 35 CP_1823 CGAAGTCCAGGAGGAAAGCTTATGCGTAAATTAGCCCTTCCC 36 GCTGAATTCGAGCTCGGTACCTCATGGGTGGGTTTCCACAA 37 CP_1834 CGAAGTCCAGGAGGAAAGCTTATGGCATTTTTACACTTTGGCTC 38 GCTGAATTCGAGCTCGGTACCCTAGACCTTCAGTTCTGGGTGGTAA 39 CP_2613 CGAAGTCCAGGAGGAAAGCTTATGACTAACAATGTTGTTACTCCGCG 40 GCTGAATTCGAGCTCGGTACCTTATTGCTTGACAGAGAACTTGGC 41 CP_2675 CGAAGTCCAGGAGGAAAGCTTATGAAAAAGCTTCTTTGGACACTCC 42 GCTGAATTCGAGCTCGGTACCCTAAACCTGGTTAGCGGCAGC 43 CP_3087 CGAAGTCCAGGAGGAAAGCTTATGAGCACCACCACCGCG 44 GCTGAATTCGAGCTCGGTACCCTACTTCAGTAGCGGGCGGG 45 CP_3110 CGAAGTCCAGGAGGAAAGCTTATGGACATAAAGAGCATGAGCTCG 46 GCTGAATTCGAGCTCGGTACCCTACTTGTGGTGTTCCACATCCG 47 XV_130 CGAAGTCCAGGAGGAAAGCTTATGAGATCAGGAAACGCCAATC 48 GCTGAATTCGAGCTCGGTACCCTAAACCACATACCCGCGTTTG 49 XV_1076 CGAAGTCCAGGAGGAAAGCTTATGATCAGGCGGCAGCGG 50 GCTGAATTCGAGCTCGGTACCTTAGACATGTGAAATCACCCGATC 51 XV_1077 CGAAGTCCAGGAGGAAAGCTTATGTCTAATTTGATGGCATCATCG 52 GCTGAATTCGAGCTCGGTACCTTAGCGCGCAGACCACGC 53 XV_1088 CGAAGTCCAGGAGGAAAGCTTATGCTGTGGTCGATATTGGTGA 54 GCTGAATTCGAGCTCGGTACCTCATGATTGGGCCTGCGC 55 XV_1267 CGAAGTCCAGGAGGAAAGCTTATGAACACCCTGGTAAAACTTCG 56 GCTGAATTCGAGCTCGGTACCTTACCTCACATGCCTCACAATCA 57 XV_2014 CGAAGTCCAGGAGGAAAGCTTATGAGTACACCGATCAAGAACCAA 58 GCTGAATTCGAGCTCGGTACCTTACCAGAGTCCGATGACATTCAT 59 XV_2617 CGAAGTCCAGGAGGAAAGCTTATGTCGAAGCTCAAGCTCAATGA 60 GCTGAATTCGAGCTCGGTACCTTATCCGAAGCGGCCGGA 61 XV_2620 CGAAGTCCAGGAGGAAAGCTTGTGAACCTCACTCTTAAGCGCTC 62 GCTGAATTCGAGCTCGGTACCTTACTGAATTGCTTCAACTGCTGC 63 XV_2682 CGAAGTCCAGGAGGAAAGCTTGTGGATCTATCCACCTGGCTTTC 64 GCTGAATTCGAGCTCGGTACCCTAGTGAGGGTGGTGATCGTGC 65 XV_2797 CGAAGTCCAGGAGGAAAGCTTATGATTAATACCATCCGCTCCG 66 GCTGAATTCGAGCTCGGTACCTTAGGCGTCTTTCTTTTCTAGGACG 67 Cgl2103 CTATGACATGATTACGAATTCGTGCCACCACGTGGCAAA 68 ATTTTCGCAAGTACATCGAAATGCGGTGATCT 69 GCGCATAACTTCGTTGATAATGGCCCCAC 70 ACGACGGCCAGTGCCAAGCTTGGGGACTTCGAAAGCATCG 71 Ptuf + CP_103 ATGTACTTGCGAAAATCCTGTTTGATGGTG 72 CAACGAAGTTATGCGCTGAGTGGCACGAC For fluorescence quantitative RT-PCR. RT-1 ilvB TCTGAGGGACTTGGCTGTG RT-2 ACTGGGCGGTCGTTGAT RT-3 brnF CGGCGTGGCGACTTATCTCA RT-4 GGCAGGAATCCAAAGTCAGCGT RT-5 brnE TTGCGATTACAGTGGTGGCG RT-6 CCTGCTGGCATCCACATCG RT-7 lrp ATCTCGCACTGGCTGACAAGG RT-8 TGTAGCCCAAAATGATTCCTTCG RT-9 leuA AGGTTGAGTTCTCCACCGTTGTC RT-10 ATCTGCTCAACTGGTGCGGT RT-11 leuB TTGCTGTTATTGGTGGAGATGGTAT RT-12 AAGGTCATAATCGGTGGTCTCG RT-13 leuC ATGACCAGCCCCGTGGAGAAC RT-14 GTCGGGCTCGCCGTTTTCT RT-15 leuD ATCCCAGCCGTCTACCTCAAG RT-16 CGAATCGTGAGGAGAAGACAGC Supplementary Table S4. Gene annotation of L-leucine-producing strain Corynebacterium glutamicum CP.
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