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pTBR101-CM

Bifidobacterium longum–E. coli shuttle vector.

Catalog number RDB15972
Resource name pTBR101-CM
Clone info. Bifidobacterium longum–E. coli shuttle vector. Efficiently transform B. longum 105-A.
Comment For selection of E. coli transformants of this plasmid, the depositor recommends use of both Cm (2.5 microgram/mL) and Ap (100 microgram/mL) because Cm resistance conferred by this plasmid is not strong. For selection of B. longum 105-A transformants, the depositor uses Cm at a concentration of 2.5 microgram/mL. They can successfully select true transformants at this concentration, but it depends on the Bifidobacterium strain. If endogenous Cm resistance is higher than 2.5 microgram/mL in your strain, the depositor recommends change of the CmR marker to the other appropriate markers that you used might be necessary. CmR region can be easily removed from pTBR101-CM by HindIII digestion. Please refer Hirayama et al., AEM, 78, 4984-4994, 2012 for further information. B. longum 105-A (JCM 31944) is available from JCM as a host strain.
Vector backbone pTBR101-CM (Plasmid)
Size of vector backbone 8.4 kb
Selectable markers Cm^r, Amp^r
Reference of insert sequence AB688000 (DDBJ accession number)
Depositor|Developer Fukiya, Satoru | Kano, Yasunobu |
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External Database

            Reference sequence
              

            Distribution information

            Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
            Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested. (Hirayama et al., Appl. Environ. Microbiol., 78(14):4984-4994 (2012)). The availability of the BIOLOGICAL RESOURCE is limited to a not-for-profit academic purpose.
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            提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Hirayama et al., Appl. Environ. Microbiol., 78(14):4984-4994 (2012))。本リソースは,非営利の学術研究に限って提供する。
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            RDB15972 pTBR101-CM DNA solution

            Please review the QC test results indicated by check icon below as well as clone information before placing your order.

            References and tips

            Electronic file

            Original reference

            original Hirayama, Y., Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functional analysis of the α-galactosidase gene for raffinose assimilation. Appl. Environ. Microbiol. 78 (14): 4984-4994 (2012). PMID 22582061.

            Further references such as user reports and related articles (go to bottom)

            Featured content

            Featured content Bacteria host strains (en) (English text)
            Featured content Bacteria host strains (ja) (Japanese text)

            Sequence information

            RIKEN BRC has sequenced portions of this material for quality test.
            Please review the QC test results indicated by check icon as well as clone information before placing your order.

            Test sheet RDB15972_A8Fbp1-2.pdf check

            Nucleotide sequence of a portion of this resource (if available).

            Primer: pBS423_seq3
            Region: rep A 3',rep B
            Sequence file: RDB15972_A8Fba.seq check
            >D04829A1-A8Fb_5_pBS423_seq3_A01_01_ABI08.ab1
                1 TGGGGCCGCC GGGGTGAAAA GGCTTCGTTT GTTGACGTTG ATTCGTCTGA GGGCCGTTTA
               61 ATGAGCCTGT GGGGGCATCG GGACATGCCG CCCAACTTCA TCGCGGAGAA TCCGGCCAAC
              121 GGCCACTGCC ACGCGGGCTG GGTGCTCACC GAACCCGTCT GCCGCACCGA CATGGCCAGA
              181 CTCAAGCCCC TGAAGCTCCT GCACGCCGTC ACCGAGGGGC TGCGCCGCAG CGTGGACGGC
              241 GACGAGGGCT ATTCCGGCCT GCTCATGAAG AACCCGTTGA GCGACGCGTG GGACAGCGAC
              301 CTGTGCCGCG AGGACACCTA CGACCTGCCC GACCTCGTGG CCGCGCTCGA AGCGCACGGG
              361 GACATGCCGC CCAAGAGCTG GACGCGCACC AAACGCGCCC GCGAGGTCGG AGTGGGCCGC
              421 AACTGCACCC TGTTCGACGA GGCCCGCACC CTCGCCTACC GCGAGGTGCG CCGACTGCCT
              481 GACCGCACGC CCGCCTCCTC CGACCTGCTG CGCGAGTACG TGCGCCGCAC CTGCCACGAA
              541 ATCAACGCCT CGTTCCCCGA CCCGCTGCCC GTGCGCGAGG TCAACGACAC CGCCAAGAGC
              601 ATCCACAAGT GGATCACCAC GCGAAGCCGC ATGTGGCGAG ACGGTGCCGT CGCCAACGCC
              661 GCCACGTTCG TCGCCATCCA ATCCGCACGC GGAAAGAAAT CGGGCGAGGC CAGACAAAAC
              721 GCATTTGAAG AGAAGTTCGC GCAGTACGCG CAGGAGGTGC TTGGACAATG ACCATCCAGA
              781 CAATTCGCAA GAAGCGCCCA TTGCCCGCCA AGGAGTTAGC CGAGGCCTAT GGGGTGTCCG
              841 TGCGCACGAT CAAGTACTGG AATTCGCAAA CGCGCGAGGG ACTGGATAGA CGAGCAGGCA
              901 ACGTTGCGCG AATCCATCCG CGCCTACCAC GACGATGACG GGCCACTCGT GGAGCCAGAC
              961 CGCCGAACAC TTCAACATGA CCCAAGGTGC CGTGCGCCAA CGCGCCTACC GCGGCCCGCA
             1021 AGGGAAACGC CGGAGGTCGA GGCCAAAAGC TCGCAAGGGC CCGGGAAGTA GCAAAGTGGA
             1081 CGTGAACGTT ATATATTACG TTCAAACTTC GGATTTCGGC GGGGTGCCGG AAGTCGGCTT
            1141 GGCCCGGGGG AAAAAGAATG C
            //
            Primer: pTBR101CM_seq7
            Region: rep B,pTB4_P2,pTB4_p3
            Sequence file: RDB15972_A8Fbb.seq check
            >D04829A1-5_A8Fb_pTBR101CM_seq7_B01_02.ab1
                1 TGAAAGGGGC GGCCGAATCC ATCCGCGCCT ACCACGACGA TGACGGCCAC TCGTGGAGCC
               61 AGACCGCCGA ACACTTCACC ATGACCCAAG GTGCCGTGCG CCAACGCGCC TACCGCGCCC
              121 GCAAGGAACG CGAGGCCGAG GCAAAAGCCG CAAGGCCGGA GTAGCAAGTG ACGTGAACGT
              181 ATATACGTTC ACTCGATTCG CGGTGCGAAT CGGCTTGCCG GGAAGACCCA TGCCCCATAG
              241 TCGGCAGAGC CTCCAGACCG CTCACGCGGC ATCGTGCCAC GACTGTGGCA CATCACGTAA
              301 CACCCGCACA TCCTCACGGT GGTGCGCGGG TGTTACGTGA TTCGAACCAT CGGCCTGACA
              361 GGTCTCAGAT CGCTCCAGCG AGCTTACGGC GCATGTACTC GCTGCGGCTG ATGTGGTACC
              421 GCCTGGCCAT CGCGGTCAGA CCGTCCAGCG TGCTCTTGGG CATGCGGACG GACACGCTGA
              481 CCATCTGCTC GTCGGGCTCG GCGAGATGGA GCCCGTAGTA CACGCGCCCC GTCAGGTCGT
              541 CCGGGACAGT CTCGGACTCG GCCATCCGCT CATCCTCCAG CACCTGCTCC TCGGTCAGCC
              601 CGACCAGCGC CATCAACTGC CTGTCGCGTT CGCTGAAATC ACTCATAGCG TCCTCCCATC
              661 GCCGCGCAGA CGGTCTATCT CGCGCCTGAA CTTCTTGGTC GGCGGCGTGA ACGCGTGGTA
              721 GATCACCACG AGGTCGCCGA CGACCCTGTA CAGCATCTCG ACGTTCCTGC CGCGCCCGTC
              781 GAGCCCTATC GCCATCCACG CGCCGGACTC GCGCTCCGCG TCCACCATGA CGCTGCGGAA
              841 CGCGGTCACC ACGTCCTTCT CGGACAGTTC TGGGTGACGC TCCATCACCC TGTCAAAGCA
              901 CCTGGGATCC TCTACGCCGG ACGCATCGTG GCCGGCATCA CCGGCGCCAC AGGTGCGGTT
              961 GCTGGCGCCT ATATCGCCGA ACATCACCGA TGGGGGAAAG AATCGGCTTC GCCACTTCGG
             1021 CTTCAATGGA GCGGCTTTGT TTCGGCGTGG GTATTGGGTG GTCAGCCTCG TGGCCCGGGG
            1081 AACTTGATGG GCGCCAATCA TCGTTGCAAT GCAACCCAAT TCCTTTTGTC G
            //
            Primer: pTBR101CM_seq3
            Region: pTB4_p1,repA 5'
            Sequence file: RDB15972_A8Fbc.seq check
            >D04829A1-A8Fb_5_pTBR101CM_seq3_B01_02_ABI08.ab1
                1 TTGCTTGTGT TAATCCCCCG CGGAGGCGGT CTGGAGGCTC CGCCGGACCA TGGGGCATGG
               61 GTCTTCCCGG CAAGCCGATT CGCACCGCGA ATCGAGTGAT TGTAGCTACA ATCACGTCGC
              121 TTGCTACTCC GGCCTTCGGC ACTCGGGTTG TTGCTCCTGC GCCGAAGGGC GAGGGAGGCT
              181 GCGGGCTCGG TTCATGTACA TGAACCGTCT AGCTTGCTTA CCTTCGATTT GATGGACATC
              241 TCCATGTGGA TGTCCATGGA GATGTCCATG GAGATGTCCA TGGAGATGTC CATGGAGATG
              301 TCCATGGAGA TGTCCATGTG GGTGTCCATG TGGGTGTCCA TGGATATATC CATCTGAGCG
              361 CATAGGAAAA CGCATGAGGG TGGATGATCG TTCGGGGTGA TGCCTTCTTC CACGAACGGG
              421 AAGGGGCTCT TCCTTTTTCT GGAATGTCAT CTTCCAGCGC CCTTCGCCTT GAGCTCCTTG
              481 AGCAGCATCG TCGCGTTGTC GCGGATGCTC GCGTCCGTGA GCGGGGAGAG CTCGTCCGCA
              541 TAGGAGCGGC AGAGGAGCGC GTACACCCGT TCCCGCACCC AGGCGCTGCG CGCGATGCCC
              601 AGCTTGAACG CGGCCCGGCA CAGCATCTCG TATTCGTCGC CGGTGAGCAG GACATTGAGC
              661 TGGCGTCAAA AGTTGATTAG TCATGCCGTG TATATACGTC CGGGGTGTTT TCGGCCAGAT
              721 TCCAACAGTT TTGGGCCACT CCGGGAAGGA ACAGCCGACG CTTCGCTGGG CTGAACCATC
              781 TGACTTGGTT CGGCGCACGC TTGTGCAATG CACAAATGTG CAATAGGATG TGAGCTATGA
              841 GCAACGAAAT ATGTGCAGGG GGACGCTGGA GCTGACGCGG GCGTTCGACG GGCTGGGTGG
              901 CTTCCCCAGC GTCCCCTGTG GCTGCGACGA TGACTACTCC CAGCTGGGTG CGGCCGGAAG
              961 CCGCATCGAA CGCGGCTCAG AATGGCAAGG CACATCGAAG GCGAAATCCG GCCAGCCGCA
             1021 AGGTGGAACA TTGCTCCGTT GTTTGGAACG TTGGATTTCG CTCTTGAGGG CCCGTTTAAA
             1081 TGGAAGCCCT TGTGGGGGGC CATCCGGGGA ACATGTGCCC GGCCCCAAAC CTTTTCCATC
             1141 CGCGGAGAAA TCCGGGCCCA ACGGGGCCAC CTGTGTCCCC ACGCCCGCCG GGGCCGATGT
            1201 GGGAGTTGG
            //
            Primer: Amp_R
            Region: Amp R pro,Cm R
            Sequence file: RDB15972_A8Fbd.seq check
            >D04829A1-A8Fb_5_Amp_R_D01_04_ABI08.ab1
                1 CCCCTTTTTT TAATAATTAT TGAGCATTTA TCAGGGTTAA TTGTCTCATG AGCGGATACA
               61 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG
              121 TGCCACCTGA CGTCTAAGAA ACCATTATTA TCATGACATT AACCTATAAA AATAGGCGTA
              181 TCACGAGGCC CTTTCGTCTT CAAGAATTCT CATGTTTGAC AGCTTATCAT CGATAAGCTT
              241 GGATCTGGAG CTGTAATATA AAAACCTTCT TCAACTAACG GGGCAGGTTA GTGACATTAG
              301 AAAACCGACT GTAAAAAGTA CAGTCGGCAT TATCTCATAT TATAAAAGCC AGTCATTAGG
              361 CCTATCTGAC AATTCCTGAA TAGAGTTCAT AAACAATCCT GCATGATAAC CATCACAAAC
              421 AGAATGATGT ACCTGTAAAG ATAGCGGTAA ATATATTGAA TTACCTTTAT TAATGAATTT
              481 TCCTGCTGTA ATAATGGGTA GAAGGTAATT ACTATTATTA TTGATATTTA AGTTAAACCC
              541 AGTAAATGAA GTCCATGGAA TAATAGAAAG AGAAAAAGCA TTTTCAGGTA TAGGTGTTTT
              601 GGGAAACAAT TTCCCCGAAC CATTATATTT CTCTACATCA GAAAGGTATA AATCATAAAA
              661 CTCTTTGAAG TCATTCTTTA CAGGAGTCCA AATACCAGAG AATGTTTTAG ATACACCATC
              721 AAAAATTGTA TAAAGTGGCT CTAACTTATC CCAATAACCT AACTCTCCGT CGCTATTGTA
              781 ACCAGTTCTA AAAGCTGTAT TTGAGTTTAT CACCCTTGTC ACTAAGAAAA TAAATGCAGG
              841 GTAAAATTTA TATCCTTCTT GTTTTATGTT TCGGGTATAA AACACTAATA TCAATTTTCT
              901 GTGGTTATAC TAAAAGTCGT TTGTTGGGTT CAAATAATGA TTAAATATCT CTTTTCTCTT
              961 CCAATTTGTC TAAATCAATT TTATTAAAGG TTCATTTGAT ATGCCTTCCT AAATTTTTAT
             1021 CTAAAGTGAA ATTAGGAGGC TTACTTTGTC TGCTTTCTTC ATTAGAATCA ATCCTTTTTT
             1081 AAAAGTCATA TTAACTGGTA ACGTAAATAT GTATTTTGAA AAATAATCCC AACTTTTTAT
            1141 CCCAATTTTT TCGGTAATG
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Hirayama, Y., Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functional analysis of the α-galactosidase gene for raffinose assimilation. Appl. Environ. Microbiol. 78 (14): 4984-4994 (2012). PMID 22582061.
            reference Nishiyama, K., Bifidobacterium bifidum Extracellular Sialidase Enhances Adhesion to the Mucosal Surface and Supports Carbohydrate Assimilation. MBio 8 (5). pii: e00928-17 (2017). PMID 28974612.
            reference Fukiya, S., Development of practical gene-mutagenesis systems in bifidobacteria. (Japanese text) Seibutsu-kogaku 94 (3): 110-116 (2016).

            2022.05.17

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