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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.
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 nucleotide sequence AB688000 (DDBJ accession number)
Depositor Fukiya, Satoru | Kano, Yasunobu |

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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RDB15972 pTBR101-CM DNA solution

Ordering Information [in Japanese] [in English]

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. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB15972_A8Fbp1.pdf

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

Primer: pBS423_seq3 | Sequence: RDB15972_A8Fba.seq
>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: pTB101CM_seq7 | Sequence: RDB15972_A8Fbb.seq
>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: pTB101CM_seq3 | Sequence: RDB15972_A8Fbc.seq
>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 | Sequence: RDB15972_A8Fbd.seq
>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).

2019.11.02

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